A5088062100- Blood Coagulation and Thrombosis Mechanisms
- Platelet Disorders and Treatments
- Endoplasmic Reticulum Stress and Disease
- Blood properties and coagulation
- Protease and Inhibitor Mechanisms
- Cancer, Hypoxia, and Metabolism
- Angiogenesis and VEGF in Cancer
- Retinoids in leukemia and cellular processes
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Cell Adhesion Molecules Research
- Complement system in diseases
- ATP Synthase and ATPases Research
- Redox biology and oxidative stress
- Mitochondrial Function and Pathology
- Hemophilia Treatment and Research
- Monoclonal and Polyclonal Antibodies Research
- Biotin and Related Studies
- Protein purification and stability
- Adenosine and Purinergic Signaling
- Protein Structure and Dynamics
- Systemic Lupus Erythematosus Research
- Hemoglobin structure and function
- Cellular transport and secretion
- Protein Interaction Studies and Fluorescence Analysis
- Nanoplatforms for cancer theranostics
Centenary Institute
2016-2025
The University of Sydney
2016-2025
University of Technology Sydney
2025
Australian Cancer Research Foundation
2022
The Heart Research Institute
2022
National Health and Medical Research Council
2015-2021
Cooperative Trials Group for Neuro-Oncology
2019
University of Salford
2007-2017
UNSW Sydney
2007-2016
Cancer Institute of New South Wales
2010-2015
Fibrin II monomer has a dramatic inhibitory effect on the rate of heparin-catalyzed inactivation human alpha-thrombin by antithrombin III. At 6 microM fibrin monomer, equivalent to concentration fibrinogen in plasma, second-order constant was reduced factor 308--from 2.05 x 10(8) M-1.s-1 6.65 10(5) M-1.s-1. minimally affected uncatalyzed thrombin showing reduction only 1.6. Fibrinogen and product plasmin degradation fibrinogen, fragment E, at concentrations also decreased for inactivation,...
Cell-surface tissue factor (TF) binds the serine protease VIIa to activate coagulation or, alternatively, trigger signaling through G protein-coupled, protease-activated receptor 2 (PAR2) relevant inflammation and angiogenesis. Here we demonstrate that TF.VIIa-mediated cell involve distinct cellular pools of TF. The surface-accessible, extracellular Cys186-Cys209 disulfide bond TF is critical for coagulation, protein isomerase (PDI) disables by targeting this disulfide. A mutant (TF C209A)...
Disulfide bonds have been generally considered to be either structural or catalytic. Structural stabilize a protein, while catalytic mediate thiol−disulfide interchange reactions in substrate proteins. There is emerging evidence for third type of disulfide bond that can control protein function by triggering conformational change when it breaks and/or forms. These thought as allosteric disulfides. To better define the properties disulfides, we analyzed geometry and dihedral strain 6874...
Morphologic assessment of lung tumors is informative but insufficient to adequately predict patient outcome. We previously identified transcriptional profiles that survival, and here we identify proteins associated with survival in adenocarcinoma. A total 682 individual protein spots were quantified 90 adenocarcinomas by using quantitative two-dimensional polyacrylamide gel electrophoresis analysis. leave-one-out cross-validation procedure the top 20 survival-associated Cox modeling...
Tissue Factor (TF) is the mammalian plasma membrane cofactor responsible for initiation of blood coagulation. Binding coagulation factor VIIa to TF activates serine proteinase zymogens factors IX and X by limited proteolysis leading formation a thrombin fibrin meshwork that stabilizes thrombus. on cells resides mostly in cryptic configuration, which rapidly transforms into an active configuration response certain stimuli. The extracellular part consists two fibronectin type III domains....
Abstract The generation of an ‘angiogenic switch’ is essential for tumor growth, yet its regulation poorly understood. In this investigation, we explored the linkage between metastasis and angiogenesis through CXCL12/CXCR4 signaling. We found that CXCR4 regulates expression secretion glycolytic enzyme phosphoglycerate kinase 1 (PGK1). Overexpression PGK1 reduced vascular endothelial growth factor interleukin-8 increased angiostatin. At metastatic sites, however, high levels CXCL12 signaling...
Abstract Protein oxidation sits at the intersection of multiple signalling pathways, yet magnitude and extent crosstalk between other post-translational modifications remains unclear. Here, we delineate global changes in adipocyte networks following acute oxidative stress reveal considerable cysteine phosphorylation-based signalling. Oxidation key regulatory kinases, including Akt, mTOR AMPK influences fidelity rather than their absolute activation state, highlighting an unappreciated...
Abstract Protein disulfide isomerase (PDI) is an oxidoreductase essential for folding proteins in the endoplasmic reticulum. The domain structure of PDI a – b b′ x a′ , wherein thioredoxin-like and domains mediate bond shuffling are substrate binding. connected via x-linker, 19-amino-acid flexible peptide. Here we identify class compounds, termed bepristats, that target substrate-binding pocket . Bepristats reversibly block binding inhibit platelet aggregation thrombus formation vivo...
Force-dependent binding of platelet glycoprotein Ib (GPIb) receptors to plasma von Willebrand factor (VWF) plays a key role in hemostasis and thrombosis. Previous studies have suggested that VWF activation requires force-induced exposure the GPIb site A1 domain is autoinhibited by neighboring A2 domain. However, biochemical basis this "mechanopresentation" remains elusive. From combination protein chemical, biophysical, functional studies, we find autoinhibition controlled redox state an...
Platelet-neutrophil interactions contribute to vascular occlusion and tissue damage in thromboinflammatory disease. Platelet glycoprotein Ibα (GPIbα), a key receptor for the cell-cell interaction, is believed be constitutively active ligand binding. Here, we established role of platelet-derived protein disulfide isomerase (PDI) reducing allosteric bonds GPIbα enhancing ligand-binding activity under conditions.Bioinformatic analysis identified 2 potential GPIbα. Agglutination assays, flow...
Protein disulfide isomerase (PDI) facilitates proper folding and bonding of nascent proteins in the endoplasmic reticulum is secreted by cells associates with cell surface. We examined consequence over- or underexpression PDI HT1080 fibrosarcoma for redox state cell-surface protein thiols/disulfides. Overexpression resulted 3.6-4. 2-fold enhanced secretion 1.5-1.7-fold increase surface-bound PDI. Antisense-mediated caused 38-53% decreased 10-33% decrease Using 5,5'-dithio-bis(2-nitrobenzoic...
N-dansylhomocysteine (DnsHCys) is quenched on S-nitrosation. The product of this reaction, N-dansyl-S-nitrosohomocysteine, a sensitive, direct fluorogenic substrate for the denitrosation activity protein disulfide isomerase (PDI) with an apparent K(M) 2 microM. S-nitroso-BSA (BSA-NO) competitively inhibited reaction K(I) 1 oxidized form DnsHCys, N,N-didansylhomocystine, rapidly accumulated in cells and was reduced to DnsHCys. fluorescence DnsHCys-preloaded human umbilical endothelial hamster...
Extracellular manipulation of protein disulfide bonds has been implied in diverse biological processes, including penetration viruses and endotoxin into cells activation certain cytokine receptors. We now demonstrate reduction one or more the serine proteinase, plasmin, by a reductase secreted Chinese hamster ovary HT1080 cells. Reduction plasmin bond(s) triggered proteolysis enzyme, generating fragments with domain structure angiogenesis inhibitor, angiostatin. Two known reductases cultured...
Plasma von Willebrand factor (vWF) is a multimeric protein that mediates adhesion of platelets to sites vascular injury. Only the very large vWF multimers are effective in promoting platelet flowing blood. A disulfide bond reductase plasma reduces average multimer size secreted by endothelial cells. This activity has been isolated from human cell conditioned medium and shown be trimeric glycoprotein, thrombospondin-1 (TSP-1). Incubation purified TSP-1 with resulted formation thiol-dependent...
Tumor necrosis factor α-converting enzyme (TACE) is responsible for the shedding of cell surface TNF. Studies suggest that reactive oxygen species (ROS) mediate up-regulation TACE activity by direct oxidization or modification protein. However, these investigations have been largely based upon nonphysiological stimulation promonocytic lines which may respond and process differently from primary cells. Furthermore, investigators relied substrate as a surrogate quantification. We addressed...