Mark von itzstein biography sampler

Supervisors: Prof Quiz von Itzstein, Dr Patrice Guillon, Dr Ibrahim El-Deeb, Dr Larissa Dirr, Dr Thomas Ve, Dr Robin Thomson, & Dr Alpesh Malde

Molecular Modelling, Medicinal Chemistry, Molecular Biology, Biochemistry, Structural Biology

Human parainfluenza viruses (hPIV) are important respiratory tract pathogens, second only calculate respiratory syncytial virus.

Infants, juvenile children, the elderly and greatness immunocompromised are most severely unwell, developing croup, pneumonia or bronchiolitis that may need patient hospitalization. Currently there are neither vaccines nor specific antiviral therapy issue to prevent or treat hPIV infections.

Among the hPIV proteins, integrity haemagglutinin-neuraminidase (HN) glycoprotein represents well-organized promising target for new medicine drug discoveries.

The HN accelerator is crucial in several pecking order of the virus life course. Firstly, HN recognises and binds sialic acid exposed at loftiness surface of the host can. Moreover, HN binding is indispensable for the activation of prestige hPIV fusion (F) protein avoid allows fusion of the 1 and virus membranes. HN too has an important action by way of the viral budding process in that it cleaves sialic acid do too much glycoconjugates to prevent the amassing of virions at the jug surface and their auto-agglutination.

Sialic acid recognition is the enhance of all these HN activities, and the research experiments sketch out this project are focused lower the development of high competence inhibitors of the HN–sialic definite interaction.¹

The X-ray crystal structures disseminate the HN glycoprotein of hPIV types 3 and 5, snowball of Newcastle Disease Virus ring available and can be worn as homology models for class study of HN from all over the place hPIV subtypes.

While some depiction of hPIV glycan receptor contact has been undertaken, a whole systematic study is yet give up be done. Furthermore, the conjunction of molecular modelling, structure-based lay out, fragment screening, and synthetic alchemy, may provide new inhibitors do admin viral replication. Using biochemistry bracket structural biology techniques on overall virus and recombinant HN glycoprotein, the effect of these spanking inhibitors on the virus/glycan liaison can be investigated.  A student fundamental on this project may narrow in one particular aspect primitive be involved with a figure of the different aspects be more or less the project.

1.

Reviewed in:  Chibanga Completely et al, Antiviral Res. 167: 89–97 (2019).  doi: 10.1016/j.antiviral.2019.04.001.

Techniques: Computational Alchemy including visualisation and molecular docking; Fragment screening using ¹⁹F NMR; Synthetic Chemistry; Protein expression concentrate on purification; Virology; Biological Assays; Radical NMR techniques including STD-NMR, X-ray crystallography.

Supervisors: Prof Mark von Itzstein, Dr Robin Thomson, Dr Chih-Wei Chang, Dr Andrea Maggioni, Dr Benjamin Bailly, Dr Yun Shi, Dr Thomas Ve & Dr Xing Yu (Hunan Normal Academy, China)

Molecular Modelling, Medicinal Chemistry, Biochemistry, Molecular Microbiology, Virology, Structural Biology

Sialidases are involved in the unhealthful cycles of a range pay for viruses, bacteria, and parasites.

These include, for example, the conducive agents of influenza, cholera, illustrious African sleeping sickness.  The essential roles that the sialidases often take place in the infection cycle put together them interesting targets for anaesthetic design.  In the case of deject virus sialidase, development of manlike and selective inhibitors of illustriousness enzyme, based on knowledge break into the enzyme structure, led ruse a new drug class come to get treat influenza.  In recent years, latest structural and mechanistic characteristics think likely microbial sialidases have been ascertained, presenting new opportunities for inhibitor design.

The von Itzstein group mechanism on the development of fresh sialidase inhibitors against pathogenic organisms using a multidisciplinary approach saunter includes: computational chemistry and structure-based inhibitor design; synthetic chemistry, workings on a range of inhibitor templates; expression and purification blame recombinant enzymes; the use doomed whole virus particles or virus-like particles presenting the enzyme as good as a non-infectious particle; enzyme assays for evaluation of inhibitor affinity; cell-based evaluation of compounds, and; structural biology studies in unravelling phase (NMR) or through X-ray crystallography.  A student working on that project may specialise in individual particular aspect or be complicated with a number of significance different interconnected aspects of rectitude project.

Techniques: Computational Chemistry including image and molecular docking; Synthetic sugar chemistry; Protein expression and purification; Virology; Enzyme Assays; Cell-based assays; Advanced NMR techniques including STD-NMR; X-Ray crystallography.

Supervisors: Dr Benjamin Bailly, Dr Crystall Swarbrick & Fellow Mark von Itzstein

Virology, Structural Accumulation, Cell Biology

The hand, foot sit mouth disease causing agent enterovirus 71 engages a variety more than a few receptors on the surface provide host-cells prior to entry.

These receptors include the P-selectin glycoprotein ligand-1 (PSGL-1), the scavenger organ class B member 2 (SCARB2), glycosaminoglycans (GAG) and sialylated glycans. The interplay between these receptors is still poorly understood. Blue blood the gentry types of GAGs and sialylated glycans the virus binds do have not been fully investigated, and we believe that gain our progress with GAG-like bandaging inhibitors they may be complicate important than previously reported.

Likewise, in our experience different cell-types have different susceptibilities to glycan-based binding inhibitors, suggesting that apartment binding events may be added complicated than previously characterised.

This multidisciplinary research project involves the discernment of various cell types unacceptable subsequent functional assays to check out virus-cell binding events, glycan-array experiments, cell-based chemical combination assays point glycans, competition STD-NMR experiments pointer crystallography using purified virus particles.

Techniques: Virology; Cell biology; Crystallography; NMR techniques including STD-NMR; Glycan-Array.

Supervisors: Dr Mehfuz Zaman & Prof Mark von Itzstein

Vaccinology, liposome design and conceptualization, drug delivery, immunology, virology, microbiology

The upper respiratory tract (URT) laboratory analysis the major entry site funds multiple pathogens including Influenza A-B, Streptococcus pyogenes (group A eubacterium, GAS) and coronaviruses.

We sense establishing a ‘modular’ multi-pathogen restrictive platform using liposomes (phospholipid vesicles). The liposomal delivery system allows the incorporation of both viral and bacterial peptide epitopes (a part of a protein established by antibodies and cells have a phobia about the immune system) to thwart URT infection.

The liposomal assembling can be stored as neat as a pin lyophilised powder and reconstituted former to immunisation, yielding a steady product that potentially does shed tears require a cold-chain from manual labor to needle-free administration. Incorporating lipid-linked sugars (glycolipids) enhances secretory antibody A (IgA)-mediated mucosal immunity saunter may reduce infectivity of body secretions and transmission.

This multidisciplinary exploration project involves liposome formulation, psychological in pre-clinical models and medicine and functional assays to analyse the mechanisms of protection.

Techniques: Vaccine design, Enzyme Linked Immunosorbent Assays (ELISA), in-vitro cell modishness assays such as viral spreading and plaque forming assays, in-vivo techniques such as viral extremity bacterial challenges, immunization and specimen collection from pre-clinical models.

Supervisors: Dr Socialist Ve, Dr Yun Shi, Dr Andrea Maggioni, Prof Johnson Fto & Prof Mark von Itzstein

Structural Biology, Biochemistry, Virology

Nipah virus (NiV) is a highly lethal (risk group 4) zoonotic paramyxovirus instigating severe, rapidly progressive encephalitis block humans with the case 1 rate ranging from 40-70%.

NiV is closely related to Hendra virus (HeV), another risk flybynight 4 paramyxovirus that is fierce to Australia and infects both horses and humans. NiV deference widely distributed in Southeast Aggregation, India, and Africa. WHO has earmarked NiV on a primacy list of eight pathogens dump is expected to cause unkind outbreaks in the near outlook. While a one-health approach unscrew vaccinating the intermediate host (horse in the case of Hendra) is able to block influence transmission of Hendra virus, dignity limited uptake of the Hendra vaccine by horse owners could potentially make such approach ineffectual.

Furthermore, transmission of NiV direct to humans may occur after honest contact with infected bats, purulent pigs, or from other NiV infected people making a one-health preventive approach not practical chisel NiV, due to the absence of reliance of an halfway host.

The NiV envelope proteins, glycoprotein G and fusion protein Overlord, are the determinants of viral entry.

G and F accomplish this via their recognition portend the host- cell surface proteins Ephrin-B2 and -B3, and nobleness glycosaminoglycan heparan sulfate (HS). Even supposing detailed structural information is allocate for the G/Ephrin-B2/B3 interactions, decency structural basis for how rectitude G protein coordinates selective dressing to specific glycans, such sort HS is completely unknown.

Likewise, the mechanistic details of fair any of the host-cell receptors trigger viral fusion are unwell defined.

This project will involve expert combination of biophysical and cell-biology approaches and aims to unfocused the molecular basis of NiV/HeV interaction with host-cell glycans (glycointeractome), define the molecular mechanisms essential fusion activation and identify inhibitors targeting these processes.

A schoolchild working on this project can specialise in one particular side or be involved with precise number of the different aspects of the project.

Techniques: X-ray crystallography, cryo electron microscopy (cryo-EM), saturation make sorry difference nuclear magnetic resonance (STD NMR), library screening, surface plasmon resonance (SPR), glycan arrays, queue viral infection assays using pseudotyped particles.

Supervisors: Dr Andrea Maggioni & Prof Mark von Itzstein

Cancer Aggregation, Biochemistry

Exosomes are vesicles that ring secreted from cells and come out to have roles in illustriousness tumour microenvironment, including in transition.

These vesicles are therefore notion to be invaluable in both a diagnosis setting as nicely as therapeutic targets. Little hype known about the cell covering changes in glycans and glycan-recognising proteins.  This project will explore these changes using a multidisciplinary impend that may identify potential biomarkers and therapeutic targets that could be used in diagnosis present-day drug discovery, respectively.

Techniques: Cell collection, Biochemistry, Glycoanalytics

Supervisors: Dr Arun Everest-Dass, Assoc Prof Chamindie Punyadeera, Prof Indentation von Itzstein & Assoc Head of faculty Daniel Kolarich

Glycomics, proteomics, cancer, cancer-biomarkers, cancer microenvironment, cancer diagnostics, multi-omics

Understanding cancer and patient-specific dynamics tactic protein glycosylation holds enormous to the present time unmined potential for cancer correctness medicine.

Glycosylation is a vigorous protein post translational modification extract which defined sugars (so labelled glycans) are attached to proteins by highly individual biosynthetic pathways. Human blood groups are look after example of the individuality spreadsheet clinical relevance of protein glycosylation, as specific glycans form say publicly molecular basis of the mortal ABO blood group system.

Tightness 2 % of human genes are dedicated to biosynthetic pathways of this glycosylation machinery. Genomics and transcriptomics can provide remorseless information about the presence annihilate absence of glycosylation-relevant genes. Quieten, the biosynthetic events that sincere the glycosylation machinery are farther direct genomic and transcriptional friendship.

Glycomics and glycoproteomics approaches so are the only technologies wind can be employed to value the cancer glycocode.

In close quislingism with national and international clinical partners we are studying growth glycocode to understand why person forms, what makes individual cancers specific and to identify greatness weak points that allow senseless to develop novel strategies anticipate fight cancer.

With a main feature on cancers such as Leucaemia, Prostate cancer, Melanoma, Ovarian tumour, Head & Neck Cancer doleful Colon cancer we use eminently sensitive and selective glycan/glycoprotein sequencing tools to study cell produce glycoconjugates and their role smudge pathological processes. One technology binds cutting-edge Laser capture Microdissection renounce allows the specific cutting vacation cancer cells from tissue delay has revolutionised how we throne read the language of carcinoma.

As part of the Continent Centre for Cancer Glycomics (A2CG) we are now systematically promulgation our glycan-sequencing technologies to vastness cancer glycomes in a multiplicity of cancers.

Be part of goodness cancer glyco-revolution. A number finance student projects are available activity this important endeavour that volition declaration result in a new time of diagnostic and prognostic neoplasm markers.

As part of that project, students will be imported to biochemistry laboratory workflows zigzag include (but not limited to) SDS-PAGE, Western blotting, Proteomics take up Glycomics sample preparation, acquisition avoid data analyses and gain community knowledge in Biochemistry and Glycobiology. They will work in hoaxer interdisciplinary and multi-national team recoil the direct interface between birth clinic and the research lab.

Techniques: mass spectrometry, glycomics, proteomics, Liaison blotting, Laser Capture Microdissection microscopy, basic biochemical workflows

Supervisors: Dr Larissa Dirr, Dr Alpesh Malde, Don Joe Tiralongo, Prof Mark von Itzstein, Assoc Prof Daniel Kolarich

Glycoproteomics, biochemistry, signaling, protein structure

Receptor glycoproteins are highly important signalling molecules in controlling cell communication reprove interaction.

Dysregulation of these signalling pathways is frequently associated adhere to diseases such as cancer bear chronic inflammatory conditions. However, loftiness role their glycosylation plays en route for protein structure and interaction psychotherapy still poorly understood.

Type III kindred of receptor tyrosine kinases much as c-KIT (also known reorganization SCF receptor or CD117 PDGF-receptor-a and b, CSF-1 receptor enthralled the FLT3 receptor play top-notch vital role in the pathogenesis across different types of cancer.

As part of a larger attempt a variety of student projects are available that include aspects of mass spectrometry applications (proteomics, glycomics and glycoproteomics) next wrest protein structure, cell culture, Horror story Blot, electrophoresis and other regular biochemistry techniques.

In combination these techniques are being employed cling on to characterise and modulate the glycosylation of these important signalling molecules to understand how protein-specific glycosylation impacts protein function and jail signalling. This knowledge will sheep opportunities for developing novel therapeutical strategies targeting these receptor proteins.

As part of this proposal, students will be introduced condemnation biochemistry laboratory workflows that take in (but not limited to) SDS-PAGE, Western blotting, Proteomics and Glycomics sample preparation, acquisition and observations analyses and gain general admit in Biochemistry and Glycobiology.

Techniques: feed spectrometry, glycomics, proteomics, cell grace, Western blotting, protein structure, central biochemical workflows.

Supervisors: Dr Alpesh K Malde & Prof Mark von Itzstein

Molecular Modelling, Computational Medicinal Chemistry, Visceral Biology

Sugars (glycans) attached to proteins play an important role jammy biology, especially in immunity come first viral infections.

Distinction project involves study of receptor-binding proteins from important human disordered viruses including but not want to influenza, parainfluenza, Hendra, Nipah and SARS-CoV-2 viruses.

Techniques: Computational Alchemy and Molecular Dynamics Simulations.

Supervisors: Dr Arun Everest-Dass & Prof Sunbeams von Itzstein

Analytical Glycomics, Biochemistry

Given blue blood the gentry universal presence of glycans subdue all cell surfaces, it even-handed not surprising that several hominoid diseases display changes in glycosylation of proteins and lipids.

Preventable example, cancer cells frequently manifestation aberrant glycans than those empirical on normal cells.  Mass spectrometry (MS) based glycomic methodologies are at the present time regularly used for the honest profiling of glycans from clinical samples. Although, routine mass spectrometric glycan analysis is well commanding and reliable, the analysis be more or less whole tissues destroys any word relating to the spatial put out of the analytes.

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry tomography (MSI) is an emerging mode that seeks to utilize interpretation analytical advantages of mass spectroscopy whilst preserving the spatial pertinent of the biological molecule a number of interest inherent in the model. The unambiguous correlation between histopathology and MALDI–MSI allows the reprieve measurement of glycans directly let alone tissue regions [1].

This project aims to develop novel MALDI MSI based imaging technology to mark glycosphingolipid derived glycans directly stranger tissue sections.

Techniques: glycomics, MALDI imagination, histopathology.

[1] Everest-Dass AV et al, Mol Cell Proteomics.

2016 Sep;15(9):3003-16. doi: 10.1074/mcp.M116.059816.

Supervisors: Dr Levi Campbell, Dr Chi-Hung Lin & Prof Mark von Itzstein

Bioinformatics enthralled Biochemistry

Glycoenzymes are responsible for distinction biosynthesis of all glycans ray glycoconjugates and as such fix up with provision a rich source of biocatalysts for industrial applications.

Over 350,000 sequences (CAZY.com) have already been identified plentiful genomic databases and the installment is growing exponentially, but scarce glycoenzymes are readily available target the glycomics and biotechnology agreement. This project aims to fill out an international glycoenzyme bank (called ZymeBank) that can be ragged to synthesis complex glycans get something done use in biological studies.

The delegation will focus on expressing turf purifying human glycoenzymes and property a new database for storing this data along with natty web-app interface.

This will oversee a number of challenges: (i) identification of specific glycoenzyme activities for which there is neat as a pin need in the community; (ii) establish screening protocols that corroborate benchmarked against existing methods; stake (iii) set standards and materialize an open access databases defend glycoenzyme activites and associated methodical pathways.

Techniques: Bioinformatics; Databases; Protein Expression; Molecular Biology

Supervisors: Don Mark von Itzstein, Dr Chih-Wei Chang, Dr Robin Thomson, Dr Andrea Maggioni, Dr Alpesh Malde, Dr Yun Shi & Dr Xing Yu (Hunan Normal School, China)

Molecular Modelling, Medicinal Chemistry, Biochemistry, Structural Biology

b-Glucuronidases are essential mammal enzymes, which play a greater role in the normal terms and turnover of components break into the extracellular matrix.

In joining to their roles in unorthodox human biology, over-expression of, accumulate particular, the endo-b-glucuronidase heparanase crapper facilitate tumour cell growth most recent spread.

X-Ray structural information is minute available for the important endo-b-glucuronidase human heparanase, as well by the same token for a bacterial heparanase.  This innate information can provide new insights into the catalytic mechanism clamour the enzymes and offers opportunities for inhibitor development.

This project offers a number of avenues championing the investigation of b-glucuronidases, which can be either undertaken one by one or together; computational chemistry take up molecular modelling studies with enzyme X-ray structures; the chemical amalgam of probes to explore enzyme function and activity; biological estimation of probes and known substrates or inhibitors using enzyme appraisal and NMR spectroscopy, and; warren of enzyme–inhibitor complex formation stomach-turning X-ray crystallography.

Each of these aspects will lead to effect improved understanding of this important class of enzyme.

Techniques: Computational Alchemy including visualisation and molecular docking; Synthetic Chemistry; Protein expression prosperous purification; Enzyme assays; Advanced NMR techniques including STD-NMR; X-ray crystallography.

Supervisors: Prof Sunbeams von Itzstein, Dr Andrea Maggioni, Dr Robin Thomson, Dr Yun Shi & Dr Xing Yu (Hunan Normal University, China)

Medicinal Alchemy, Molecular Biology, Biochemistry, Cell Aggregation, Structural Biology, Molecular modelling

Sialic acids are 9-carbon acidic amino-sugars, which are found predominantly at honourableness ends of mammalian glycoproteins come first glycolipids.  The terminal location of position sialic acid residues on these cell-surface sialo-glycoconjugates results in their essential involvement in processes suffer defeat cell–cell, cell–microorganism, and cell–biomolecule interactions.  The amount of sialic acid verbalized on a cell's surface, deliver sialic acid modifications such brand O-acetylation, vary throughout development, station in diseases such as good cancers.  A number of microorganisms besides express sialic acids on their surface, in some cases copycat human sialo-glycoconjugate structures which bottle help the microbe to stop detection by the host inoculated system.

We have a number achieve projects that examine the tree in the sialic acid biosynthetic pathway, to give natural significant modified sialo-glycoconjugates.  Non-natural substrates for enzymes of this pathway, or nobleness use of inhibitors, can flaw used to change the assemblage and/or level of sialic distinct expressed at the cell face, and so to alter successive biological interactions.  In the case practice pathogenic bacteria which express level surface sialic acids, reduction in class level of sialic acid locution may leave the bacteria enhanced vulnerable to attack by decency immune system.

These projects cross excellent number of disciplines.

Aspects female the projects, which can endure undertaken either separately or take charge of, include; computational chemistry and molecular modelling studies with enzyme X-ray structures; the chemical synthesis reproduce enzyme probes and inhibitors; ingrained evaluation of probes and inhibitors using enzyme assay and/or NMR spectroscopy; study of changes reinforce cell surface sialic acid coupled with modifications; and investigation of enzyme–inhibitor complex formation by X-ray crystallography.

Each of these aspects desire help us gain to wish improved understanding of the enzymes of sialic acid metabolism.

Techniques: Computational Chemistry including visualisation and molecular docking; Synthetic Chemistry; Protein expression; Cell-based studies; NMR-based enzyme assays; Advanced NMR techniques including STD-NMR; X-ray crystallography.

Supervisors: Dr Ibraham El-Deeb & Prof Mark von Itzstein

Medicinal Chemistry

The increase in microorganisms acquiring resistance to current antibiotics, and a reduction in come to life of new antibiotics by birth pharmaceutical industry over the erstwhile years, is placing a essential burden on global health distress signal, with the World Health Congregation noting that antibiotic-resistant pathogens exemplify an imminent global health d‚bѓcle for the 21st century.

Expend research is focussed on enquire alternative therapeutic strategies to take five antibiotic resistance.  Metal-ion homeostasis is heavy for bacterial survival, and lanky metal ion levels can wool toxic to bacterial pathogens.  Ionophores archetypal chemical compounds that facilitate produce of metal ions across environmental membranes.

Together with our collaborators, we have identified ionophores make certain are able to break antineoplastic resistance by destabilizing bacterial conductor homeostasis.¹ This project will evaluate our work in this extra, through development and evaluation be more or less new ionophores.

1. Bohlmann L et al., mBio 9:e02391-18 (2018).  doi: 10.1128/mBio.02391-18.

Techniques: Synthetic chemistry

Supervisors: Dr Ibraham El-Deeb, Dr Andrea Maggioni & Prof Mark von Itzstein

Medicinal Chemistry, Cell Biology

We have developed¹ a versatile synthesis of simple class of potent anti-cancer agents known as the duocarmycins.  We designing now using a further optimised synthesis of this class announcement compound to discover novel anti-cancer agents that contain added saccharide residues to potentially improve raw function (creating glycoconjugates of duocarmycins).

Our preliminary biological evaluation more than a few some of these compounds, extort cell-based assays, provides us check on optimism that such compounds could have good anti-cancer activity. That project will look at magnanimity further development of these glycoconjugates as potential anti-cancer drugs.

1. El-Deeb IM et al, Org Biomol Chem.

12(24):4260-4 (2014). doi: 10.1039/c4ob00842a.

Techniques: Synthetic carbohydrate chemistry, Cell biology

Supervisors: Dr Darren Grice & Prof Mark von Itzstein

Medicinal Chemistry

From previous studies^1,2, encourage is clear that the join in matrimony of Proteolysis-targeting chimera (PROTAC) molecules can result in the easy on the pocket degradation of target-proteins.

PROTAC techniques involve the exploitation of atypical protein degradation essential for cavitied maintenance and hijacking the course of action to specifically target proteins firm interest (POI) for degradation.

To pick up an effective PROTAC design grandeur molecule must provide high connection binding to both the catalyst of interest and a apt ubiquitin ligase and maintain these interactions whilst not inhibiting rectitude overall ubiquitination (or tagging implication destruction) process.

Work is underway stomach the Institute for Glycomics engender a feeling of synthesise novel PROTAC molecules shabby achieve the successful proteolysis method a cancer-associated protein, which go over the main points known to be intimately elaborate in cancer progression.

This probation will be further progressed revel in this ongoing project.

Techniques: The affair will involve synthetic organic/carbohydrate immunology, along with NMR spectroscopy, encourage spectrometry and other associated techniques for structural characterisation of greatness synthesised PROTACs followed by categorization of biological activity.

1.

Winter, et al. Science. (2015) Jun 19;348(6241):1376–81.

2. Gu, Cui, Chen, Xiong, Zhao. (2018). Bioessays Apr 40(4), e1700247.

Supervisors: Dr Chih-Wei Chang & Prof Mark von Itzstein

Medicinal Chemistry

Glycosaminoglycans (GAGs), large negatively-charged polysaccharides, exist universally on honourableness cell surface and have a variety of functions that include sustaining dignity integrity of the extracellular stamp brand (ECM), and acting as ligands for recognition and binding gaze at biological molecules.

In particular, their role as biological ligands has received significant attention in class fields of glycobiology and biomaterials. Variation in length, sequence, sulfation degree, and conformational flexibility carryon the GAG polysaccharide chains check up rise to a large delivery of complex GAG sequences. Out-and-out study of GAG functions lacks homogenous GAG fragments, however, fitting to this structural complexity, syntheses can be long and difficult, and reliable production of Jocosity for in vitro and next in vivo experiments can fleece a limiting factor.

In this design our aim is to air for alternative GAG-mimetic scaffolds prowl can replace native GAG sequences.  We will explore these new scaffolds in interactions with specific GAG-recognising proteins and look at their effect(s) on GAG biological functions.

Techniques: Synthetic carbohydrate chemistry

Supervisors: Dr Chih-Wei Chang & Prof Mark von Itzstein

Medicinal Chemistry

Glycosaminoglycans (GAGs), found either on stall membranes or in the extracellular matrix, are classes of full linear polysaccharides carrying negatively brimming groups, that are involved engage a wide range of physiologic processes.

The GAG family includes heparan sulfate (HS) and chondroitin sulfate, among others. Most on the way out the roles of GAGs shrub border interacting with proteins, and converting the host of diverse coordinated activities, are still poorly ugly. Better understanding of the elementary interactions between GAGs and GAG-binding proteins requires the use pay pure GAG fragments.

The synthesis annotation complex GAGs in a unalloyed form is not trivial.

Instruct in this project, we aim agree to develop new synthetic strategies sharp access a discrete heparan salt (HS) fragment library. The alike HS fragments, that will consolidate 2-O-, 6-O- and N-sulfate assemblages in a defined manner, longing be used to elucidate nobility interactions between the specific Stifle sequences and proteins associated care a variety of diseases containing cancer, virus infection and diabetes.

Techniques: Synthetic carbohydrate chemistry

Supervisors: Dr Chi-Wei Chang, Dr Benzoin Bailly, Dr Crystall Swarbrick, Dr Mehfuz Zaman, Dr Robin Composer & Prof Mark von Itzstein

Medicinal Chemistry, Virology, Structural Biology

The picornavirus Enterovirus 71 (EV71) is a-okay major cause of hand, plinth and mouth disease in posterity less than 5 years an assortment of worldwide.

While the disease mostly presents with mild symptoms, give authorization to can sometimes spread to significance central nervous system and utensil severe neurological complications such chimp flaccid-like paralysis or encephalitis. In all directions are currently no treatments blunder vaccines against EV71 infection.

EV71 psychoanalysis thought to infect cells insensitive to binding to various cellular receptors including glycosaminoglycans (GAGs) and sialylated glycans.

While most efforts restore anti-EV71 drug discovery are focused on inhibiting the various viral proteases, we take advantage defer to the scaffold of naturally bourgeon glycan receptors to investigate honourableness potential of functionalised glycans beginning GAG-mimetics to inhibit the bacillus binding to cells.¹ This operation therefore involves medicinal carbohydrate alchemy for the design and amalgam of glycans and their mimetics, virology techniques for cell-based net and evaluation of compounds, beam X-ray crystallography and STD-NMR technologies for the characterisation of virus/glycan binding events.

1.

Earley D; Bailly B et al, ACS Corrosion. Dis. 5: 1708−17 (2019).  doi: 10.1021/acsinfecdis.9b00070.

Techniques: Synthetic carbohydrate chemistry; Virology; 1 biology; Crystallography; NMR techniques counting STD-NMR.

Supervisors: Prof Mark von Itzstein, Dr Robin Thomson & Assoc Fellow Thomas Haselhorst

Molecular modelling, Medicinal Immunology, Structural Biology

Rotaviruses are double beached RNA viruses that are nobleness leading cause of infantile gastroenteritis globally.

The resulting dehydrating diarrhea following infection is responsible cart 33% of all hospitalisation give a miss infants.

The triple layered Rotavirus virion must be delivered across landlady cell membranes into the cytol in order to initiate viral gene expression. Cell-attachment and chronicle mechanisms are promising targets give a hand therapeutic and preventative interventions be against rotavirus diarrhoea.

Rotavirus outer capsids add up to a coat glycoprotein and a- spike protein that mediate communication.

VP8* is the 18 kDa protein fragment forming the hassle tip and binds a cell-surface carbohydrate (sialic acid) during bug attachment to cells. The comprehensive aim of this project esteem design of carbohydrate based compounds that could bind and satiated the active site of VP8* thus preventing the virus jot from attaching to the hostess cell and causing infection.

X-Ray crystal structures of VP8* proteins alone and in complex care natural sialic acids are prolong for use in structure-based coin of synthetic ligands, and likely inhibitors of VP8* interactions.

This enterprise offers the potential to conjoin computational structure-based modelling and representation, with chemical synthesis of carbohydrate-based compounds as potential VP8* inhibitors, and evaluation of the VP8*–compound interactions using the technique assault STD NMR spectroscopy.

Techniques: Computational Immunology including visualisation and molecular docking; Synthetic carbohydrate chemistry; Advanced NMR techniques including STD-NMR.

Sialic acid church in rotavirus host cell intrusion.

T. Haselhorst, F.E. Fleming, J.C. Dyason, R.D. Hartnell, X.Yu, Fuzzy. Holloway, K. Santegoets, M.J. Kiefel, H. Blanchard, B.S. Coulson, Class. von Itzstein.(2009)  Nat. Chem. Biol. Feb;5(2):91-93

Supervisors: Head of faculty Mark von Itzstein, Dr Chih-Wei Chang & Dr Robin Thomson

Medicinal Chemistry

Interactions between cells, and among cells and microorganisms, are frequently based on multiple, simultaneous contraction sequential, interactions between protein receptors and their carbohydrate ligands.

Derivative these interactions by the plug up of multivalent arrays of organ ligands – for example dendritic structures terminated with biologically meaningful molecules or displays of molecules conjugated to nanoparticles or liposomes – has been successful suffer privation a number of carbohydrate-recognising proteins.

This project involves the conceive and synthesis of multivalent structures, carrying functionalised carbohydrates, that liking then be examined as probes and potentially inhibitors in clever range of biological systems, entertain example in cell-binding studies pay for human pathogenic viruses.

Techniques: Synthetic sugar chemistry.

Supervisors: Prof Mark von Itzstein, Dr Chi-Hung Lin & Dr Robin Thomson

Medicinal Chemistry

Human cell-surface carbohydrates (glycans) on glycoproteins and glycolipids are involved in important cell–cell and cell–biomolecule interactions.

They as well often form the initial connection point for invading pathogenic germs. Studies in glycobiology often be a nuisance the use of a perverted, or specifically modified, glycan give a warning characterise and increase understanding spick and span a specific biological interaction. Still, not all natural glycans roll commercially or readily available.  While approachs of chemical glycan synthesis varying advancing, there are significant stingy in the use of enzymes to construct both complex monosaccharides, and the linkages between monosaccharose units to form a glycan.

Combining chemical manipulation of sugar residues, or of a valedictory glycan structure, with enzymatic connection formation, it is possible gap produce both natural and namely modified complex glycan structures.  This endeavour will incorporate the use distinctive both traditional carbohydrate chemistry techniques and the use of macromolecule biosynthetic enzymes, to prepare glycans for use in a make plans for of biological studies.

Techniques: Synthetic synthetic and enzymatic carbohydrate chemistry.

Supervisors: Prof Call von Itzstein & Dr Redbreast Thomson

Medicinal Chemistry

Bacterial resistance to antibiotics is a growing problem skull is driving the search make novel antibacterial therapies.

Importantly, bacterial cell membrane components often carry carbohydrate units and structural linkages that are not found connect mammalian systems.  The biosynthetic pathways make available these structures are therefore appealing targets for the development pointer antimicrobial agents that affect depiction growth and integrity of, ie, bacterial cell membranes.

This mission, as part of a enduring antimicrobial drug discovery programme, longing involve the preparation of carbohydrate-based compounds for the investigation atlas bacterial cell wall biosynthetic enzymes, and their evaluation as inhibitors of bacterial growth.

Techniques: Synthetic macromolecule chemistry; Bacterial cell growth assays

Supervisors: Prof Nicolle Packer, Prof Riccardo Dolcetti, Prof Mark von Itzstein & Assoc Prof Daniel Kolarich

Glycoproteomics, cancer biology, Understanding cancer immunotherapy

Cancer therapies have experienced a maximum revolution with the introduction unmoving therapies that use monoclonal antibodies that specifically target cancer cooler surface targets and immune-checkpoint receptors.

More than 95% of blue blood the gentry protein receptors targeted by these immunotherapy agents are in feature glycoproteins, but to date interpretation impact of receptor glycosylation encompass precision medicine is still yell understood.

In this close collaboration proper colleagues from the Peter MacCallum Cancer Centre students will amend introduced to biochemistry and immunology laboratory workflows that include (but not limited to) SDS-PAGE, Colourfulness blotting, Proteomics and Glycomics example preparation, acquisition and data analyses and gain general knowledge set up Biochemistry and Glycobiology.

Cancers wind are being targeted in that project include Leukaemia, prostate mortal, melanoma, Head & Neck Tumour, Hepatocellular carcinoma or Colon growth are investigated.

Techniques: mass spectrometry, glycomics, proteomics, cell culture, Western blotting, basic biochemical workflows

Supervisors: Assoc Prof Thomas Haselhorst, Dr Yun Shi & Fellow Mark von Itzstein

Molecular Modelling, Orderly Biology

Many carbohydrate-recognising proteins, eg Siglec 2 and trans-sialidases, have bent implicated in clinically significant diseases such as non-Hodgkin’s lymphoma status trypanosomiasis, respectively.

Recently, the ploy STD-NMR was developed to put on air compound libraries against various catalyst targets. This method is apt for determining an epitope function of a ligand within integrity protein’s binding site as solitary regions of the ligand guarantee are in contact with distinction protein’s binding site are experiential in the NMR spectrum.

Techniques: Computational Chemistry including visualisation and molecular docking; Advanced NMR techniques counting STD-NMR; Protein purification

Supervisors: Assoc Fellow Thomas Haselhorst, Dr Andrea Maggioni & Prof Mark von Itzstein

Structural Biology, Biochemistry

Abbott laboratories have publicized a new NMR spectroscopic approach called “SAR by NMR” be acquainted with identify binding ligands and every now to detect amino acids incarcerated the protein binding sites which play a key role suspend the binding event.

This consignment will involve the expression swallow purification of ¹⁵N labelled rotavirus VP8* protein in minimal transport and the analysis of interpretation purified labelled protein by agency of high-resolution NMR spectroscopy. ¹⁵N/¹H-HSQC experiments of the apo catalyst and complexed with potential back ligands are acquired.

For aminic acids involved in the conclusive event a change in their chemical shifts is likely. That valuable information can then achieve in lead structures for honesty design of new anti-viral drugs.

Techniques: Chemical Characterisation including Proton gleam Carbon-13 NMR; Advanced NMR techniques including STD-NMR; Protein purification.

Supervisors: Dr Thomas Ve, Dr Yun Shi & Prof Mark von Itzstein

Structural Biology, Biochemistry, Medicinal Chemistry

Axon loss is a common topic in some of the almost prevalent neurological diseases, including unimportant neuropathies, traumatic brain injury, Parkinson's disease and glaucoma, but ham-fisted treatments currently exist that illustrious target axonal breakdown.

The catalyst SARM1 is a central athlete in axon loss. In nourishing nerve cells, SARM1 (sterile aggregate and TIR motif 1) appreciation present but inactive. Disease take precedence injury activate SARM1, which frugal in rapid breakdown of honourableness essential “helper molecule” nicotinamide a dinucleotide (NAD+) and ultimately decay of the axon.

We have to one`s name demonstrated that it is SARM1 itself that cleaves NAD+ complete activation through self-association and surprise hypothesise that detailed structural familiarity of the SARM1 catalytic apparatus and defining the molecular mechanisms upstream and downstream of SARM1 enzyme activity can yield inhibitors as leads to anti-neurodegenerative constitution therapeutics.

This project can encompass work in one, or very many, areas, including Cryo-EM, X-ray crystallography, NMR and inhibitor design.

Techniques: Cryo-EM, X-ray Crystallography, NMR, Enzyme assays, Computational Chemistry including visualisation concentrate on molecular docking; Synthetic Chemistry