A5042881187- Monoclonal and Polyclonal Antibodies Research
- Protein Structure and Dynamics
- RNA and protein synthesis mechanisms
- Glycosylation and Glycoproteins Research
- Peptidase Inhibition and Analysis
- Enzyme Structure and Function
- Immune Cell Function and Interaction
- T-cell and B-cell Immunology
- Receptor Mechanisms and Signaling
- Advanced biosensing and bioanalysis techniques
- Protein Interaction Studies and Fluorescence Analysis
- Diabetes and associated disorders
- Lipid Membrane Structure and Behavior
- DNA and Nucleic Acid Chemistry
- Protein purification and stability
- Thyroid Disorders and Treatments
- Cell Adhesion Molecules Research
- Biochemical and Structural Characterization
- Immune Response and Inflammation
- Endoplasmic Reticulum Stress and Disease
- NF-κB Signaling Pathways
- Biochemical and Molecular Research
- Cellular Mechanics and Interactions
- Chemical Synthesis and Analysis
- interferon and immune responses
Monash University
2012-2025
Australian Regenerative Medicine Institute
2015-2025
MRC Laboratory of Molecular Biology
2016-2024
Discovery Institute
2018-2024
Medical Research Council
2016-2021
Victor (Japan)
2018
Abstract Optimum protein function and biochemical activity critically depends on water availability because solvent thermodynamics drive folding macromolecular interactions 1 . Reciprocally, macromolecules restrict the movement of ‘structured’ molecules within their hydration layers, reducing available ‘free’ bulk therefore total thermodynamic potential energy water, or potential. Here, concentrated solutions such as cytosol, we found that modest changes in temperature greatly affect...
Antibodies have the specificity to differentiate foreign antigens that mimic self antigens, but it remains unclear how such is acquired. In a mouse model, we generated B cells displaying an antibody cross-reacts with two related protein expressed on versus cells. cell anergy was imposed by antigen reversed upon challenge high-density antigen, leading germinal center recruitment and gene hypermutation. Single-cell analysis detected rapid selection for mutations decrease affinity slower...
Abstract Developing therapeutic antibodies is laborious and costly. Here we report a method for antibody discovery that leverages the Illumina HiSeq platform to, within 3 days, screen in order of 10 8 antibody–antigen interactions. The method, which named ‘deep screening’, involves clustering sequencing libraries, conversion DNA clusters into complementary RNA covalently linked to instrument’s flow-cell surface on same location, situ translation tethered via ribosome display, their screening...
Abstract Poor understanding of intracellular delivery and targeting hinders development nucleic acid‐based therapeutics transported by nanoparticles. Utilizing a siRNA‐targeting small molecule profiling approach with advanced imaging machine learning biological insights is generated into the mechanism lipid nanoparticle (MC3‐LNP) mRNA. This workflow termed Advanced Cellular Endocytic for Intracellular Delivery (ACE‐ID). A cell‐based assay perturbation 178 targets relevant to trafficking used...
The crystal structures of unliganded and liganded pMHC molecules provide a structural basis for TCR recognition yet they represent 'snapshots' offer limited insight into dynamics that may be important interaction T cell activation. MHC HLA-B*3501 HLA-B*3508 both bind 13 mer viral peptide (LPEP) only HLA-B*3508-LPEP induces CTL response characterised by the dominant clonetype SB27. forms tight long-lived complex with SB27, but relatively weak between HLA-B*3501-LPEP SB27 fails to trigger an...
Serine proteinase inhibitors (serpins), typically fold to a metastable native state and undergo major conformational change in order inhibit target proteases. However, lability of the serpin renders them susceptible misfolding aggregation, underlies diseases such as α1-antitrypsin deficiency. Serpin specificity towards its protease is dictated by flexible solvent exposed reactive centre loop (RCL), which forms initial interaction with during inhibition. Previous studies have attempted alter...
The enzyme glutamate decarboxylase (GAD) produces the neurotransmitter GABA, using pyridoxal-5'-phosphate (PLP). GAD exists as two isoforms, GAD65 and GAD67. Only acts a major autoantigen, frequently implicated in type 1 diabetes other autoimmune diseases. Here we characterize structure dynamics of its interaction with polyendocrine syndrome 2-associated autoantibody b96.11. Using hydrogen-deuterium exchange mass spectrometry (HDX), X-ray crystallography, cryo-electron microscopy,...
Thyroid peroxidase (TPO) catalyses the biosynthesis of thyroid hormones and is a major autoantigen in Hashimoto's disease—the most common organ-specific autoimmune disease. Epitope mapping studies have shown that response to TPO directed mainly at two surface regions on molecule: immunodominant A B (IDR-A, IDR-B). has been target for structural over 20 years; however, date, structure remains be determined. We used molecular modelling approach investigate plausible modes dimer organisation....
Recombination, the exchange of information between different genetic polymer strands, is fundamental importance in biology for genome maintenance and diversification mediated by dedicated recombinase enzymes. Here, we describe an innate capacity non-enzymatic recombination (and ligation) random-sequence oligomer pools. Specifically, examine random semi-random eicosamer (N20) pools RNA, DNA unnatural polymers ANA (arabino-), HNA (hexitol-) AtNA (altritol-nucleic acids). While DNA, proved...
Structure-function analysis of chemokines and their shared receptor may aid in the development small-molecule inhibitors.
Consensus protein design is a rapid and reliable technique for the improvement of stability, which relies on use homologous sequences. To enhance stability fibronectin type III (FN3) domain, consensus was employed using an alignment 2123 The resulting FN3 FN3con, has unprecedented with melting temperature >100°C, ΔG(D-N) 15.5 kcal mol(-1) greatly reduced unfolding rate compared wild-type. determine underlying molecular basis X-ray crystal structure FN3con determined to 2.0 Å other domains...
Significance Autoimmune type 1 diabetes is characterized by the formation of self-reactive antibodies. A prevalent human autoantigen glutamate decarboxylase (GAD)65, a highly predictive marker that can precede emergence disease up to several years. Intriguingly, closely related isoform GAD67 not immunogenic. What are determinants unique self-reactivity GAD65 vs. GAD67? We show that, unlike GAD67, flexible and exists in multiple structural forms. self-antibodies bind differentially these...
Abstract The kallikrein-related peptidase (KLK) family of proteases is involved in many aspects human health and disease. One member this family, KLK4, has been implicated cancer development metastasis. Understanding mechanisms inactivation are critical to developing selective KLK4 inhibitors. We have determined the X-ray crystal structures complex with both sunflower trypsin inhibitor-1 (SFTI-1) a rationally designed SFTI-1 derivative atomic (~1 Å) resolution, as well bound nickel. These...
The rugged folding landscapes of functional proteins puts them at risk misfolding and aggregation. Serine protease inhibitors, or serpins, are paradigms for this delicate balance between function misfolding. Serpins exist in a metastable state that undergoes major conformational change order to inhibit proteases. However, labiality the native serpin fold renders susceptible misfolding, which underlies diseases such as α1-antitrypsin deficiency. To investigate how serpins folding, we used...
Thyroid peroxidase (TPO) is a dimeric membrane-bound enzyme of thyroid follicular cells, responsible for hormone biosynthesis. TPO also common target antigen in autoimmune disease (AITD). With two active sites, an unusual enzyme, and thus there much interest understanding its structure role AITD. Homology modeling has shown to be composed different structural modules, as well propeptide sequence. During the course studies obtain homogeneous preparations recombinant studies, we investigated...
The favorable biophysical attributes of non-antibody scaffolds make them attractive alternatives to monoclonal antibodies. However, due the well-known stability-function trade-off, these gains tend be marginal after functional selection. A notable example is fibronectin Type III (FN3) domain, FNfn10, which has been previously evolved bind lysozyme with 1 pM affinity (FNfn10-α-lys), but suffers from poor thermodynamic and kinetic stability. To explore this compromise further, we grafted...
Significance Conformational diversity of foreign antigens and cross-reactivity with self are implicated in the failure to generate effective antibody responses against many challenging pathogens, but few studies directly address how these two factors affect formation. Here we this question from biophysical, structural, immunological perspectives using structurally related lysozyme proteins. The results show germinal centers have remarkable ability select producing cells along novel...
Dihydrodipicolinate synthase (DHDPS) is an essential enzyme involved in the lysine biosynthesis pathway. DHDPS from E. coli a homotetramer consisting of 'dimer dimers', with catalytic residues found at tight-dimer interface. Crystallographic and biophysical evidence suggest that dimers associate to stabilise active site configuration, mutation central dimer-dimer interface residue destabilises tetramer, thus increasing flexibility reducing efficiency substrate specificity. This has led...