A5086878223- Advanced Proteomics Techniques and Applications
- Prostate Cancer Treatment and Research
- Monoclonal and Polyclonal Antibodies Research
- Radiopharmaceutical Chemistry and Applications
- Mass Spectrometry Techniques and Applications
- Glycosylation and Glycoproteins Research
- Nanoplatforms for cancer theranostics
- Ocular Surface and Contact Lens
- Food Allergy and Anaphylaxis Research
- Genomics and Phylogenetic Studies
- Prostate Cancer Diagnosis and Treatment
- Allergic Rhinitis and Sensitization
- Salivary Gland Disorders and Functions
- Medical Imaging Techniques and Applications
- Proteoglycans and glycosaminoglycans research
- Bacterial Genetics and Biotechnology
- Bacterial biofilms and quorum sensing
- Antimicrobial Resistance in Staphylococcus
- Contact Dermatitis and Allergies
- Occupational exposure and asthma
- Molecular Biology Techniques and Applications
- Bladder and Urothelial Cancer Treatments
- Antibiotic Resistance in Bacteria
- Heavy Metal Exposure and Toxicity
- Biosensors and Analytical Detection
Parkland Health & Hospital System
2009-2023
University of Technology Sydney
2021
Public Health England
2014-2017
Macquarie University
1989-2014
Landcare Australia
2008
Texas Department of State Health Services
2006
Washington State University
2004
Walsh University
2004
University of Cambridge
2004
Australian Genome Research Facility
1997-2003
Macrophages play a key role in both normal and pathological processes involving immune inflammatory responses, to large extent through their capacity secrete wide range of biologically active molecules. To identify some these as yet not characterized molecules, we have used subtraction cloning approach designed genes expressed association with macrophage activation. One genes, designated inhibitory cytokine 1 (MIC-1), encodes protein that bears the structural characteristics transforming...
We describe the extraction and enrichment of membrane proteins for separation by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) after differential solubilization an Escherichia coli cell lysate. In a simple three-step sequential protocol applicable whole lysates, are partitioned from other cellular their insolubility in solutions conventionally used isoelectric focusing (IEF). As first step, Tris-base was to solubilize many cytosolic proteins. The resultant pellet then...
Abstract Occupancy-induced down-regulation of cell surface epidermal growth factor (EGF) receptors attenuates signal transduction. To define mechanisms through which this class occurs, we have investigated the relative roles ligand-induced internalization and recycling in process. Occupied, kinase-active EGF were internalized a high affinity, saturable endocytic system at rates up to 10-fold faster than empty receptors. In contrast, full length lacking tyrosine kinase activity underwent rate...
Abstract In this study, dithiothreitol was replaced by tributyl phosphine as the reducing agent in both sample solution for first‐dimensional isoelectric focusing and during immobilised pH gradient (IPG) equilibration procedure. Tributyl improves protein solubility focusing, which results shorter run times increased resolution. is nonionic thus does not migrate IPG, therefore maintaining conditions course of separation. The provided maintenance gives improved decreased horizontal streaking...
We have developed a quantitative method to evaluate the interaction between cell surface receptors and endocytic apparatus. This exploits occupancy-dependent changes in internalization rates that occur cells expressing high numbers of receptors. found constitutive transferrin receptor behaves as simple, first order process is unaltered by ligand. Internalization epidermal growth factor (EGF) receptor, however, saturable, second induced occupancy. EGF occurs through at least two distinct...
Circulating cancer-derived small extracellular vesicles (EVs) are nanoscale membranous shed from cancer cells that released into surrounding body fluids. Small EVs contain biomolecules associated with such as DNA and proteins for cell-to-cell communication. Therefore, have been regarded important biomarkers liquid biopsy-based diagnosis drug treatment monitoring. However, because of the high heterogeneity low level in fluids, there is a demand sensitive detection characterization at...
With the completion of many genome projects, a shift is now occurring from acquisition gene sequence to understanding role and context products within genome. The opportunistic pathogen Pseudomonas aeruginosa one organism for which available, including annotation open reading frames (ORFs). However, approximately third ORFs are as yet undefined in function. Proteomics can complement genomics, by characterising their response variety biological environmental influences. In this study we have...
Endocytic Codes in EGF Receptor19313 of these sequences the context receptor is strictly dependent on ligand binding and intrinsic tyrosine kinase activity.Because function receptors that lack self-phosphorylation sites, requirement appears due to phosphorylation a cellular component involved endocytosis. MATERIALS AND METHODSConstruction Mutant Receptors-For construction mutant receptors, vectors expressing holoreceptor c'991 c'958 f993-1022 mutants were used (15).In nomenclature used, c'...
The las and rhl quorum sensing (QS) systems regulate the expression of several genes in response to cell density changes Pseudomonas aeruginosa . Many these encode surface-associated or secreted virulence factors. Proteins from stationary phase culture supernatants were collected wild-type P. PAO1 mutants deficient one more lasRI , rhlRI vfr analysed using two-dimensional gel electrophoresis. All released significantly lower amounts protein than wild-type. Protein spot patterns each strain...
Little is known about the regulation of EGF receptor (EGF-R) trafficking following endocytosis. We investigated this by using a series EGF-R with altered cytoplasmic tails and comparing their ability to undergo recycling lysosomal targeting in both occupied empty state. found that 2-3% are internalized each minute, but rapidly recycle (t1/2 approximately 5 min). This constitutive internalization receptors was independent sequences. Occupied EGF-R, contrast, displayed much slower rate between...
Summary Hexapeptides of sequential overlapping sequences β‐lactoglobulin (BLG) were used to probe scrum from children with immediate‐type cow milk allergy for IgE binding continuous epilopes BLG in an enhanced enzyme‐linked immunosorbent assay (ELISA). Six regions identified on the molecule and these synthesized as dodecapeptides. Inhibition whole was confirm BLG‐specific each peptides. One peptide 4, showed inhibition anti‐BLG radioimmunoassay all 16 sera tested. The patterns native 4...
To assess the functional significance of phosphorylation epidermal growth factor (EGF) receptor at Thr654, we compared effects 12-O-tetradecanoyl-13-acetate (TPA) on ligand-induced internalization and down-regulation between wild-type mutant receptors that contain an alanine substitution position 654. Activation protein kinase C with TPA blocked EGF-induced Thr654 inhibited in vivo tyrosine activity by 80%. did not inhibit transferrin or constitutive EGF internalization, suggesting...
During the last decade, isolation of circulating tumour cells via blood liquid biopsy prostate cancer (PCa) has attracted significant attention as an alternative, or substitute, to conventional diagnostic tests. However, it was previously determined that localised forms PCa shed a small number into bloodstream, and large volume is required just for single test, which impractical. To address this issue, urine been used alternative truly non-invasive, patient-friendly test. end, we developed...
Abstract To understand the changes in protein expression associated with various physiological states as well development of pathological eye disease, we have begun to map components normal human reflex tears. An analytical reference tears was created using two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) pH 3.5–10 immobilized gradients (IPGs). Micropreparatively loaded gels were transferred polyvinylidene difluoride (PVDF) and analysed by a combination N ‐terminal sequence...
Proteomics is a powerful tool for analysing differences in gene expression between bacterial strains with alternate phenotypes. Staphylococcus aureus are grouped on the basis of their sensitivity to methicillin. Two-dimensional gel electrophoresis was combined MS compare protein profiles S. COL (methicillin-resistant) and 8325 (methicillin-sensitive). Reference mapping via this approach identified 377 proteins that corresponded 266 distinct ORFs. Amongst these were 14 potential virulence...
The epidermal growth factor (EGF) receptor contains multiple sites of phosphorylation on serine, threonine, and tyrosine residues. Because the biological responsiveness EGF is regulated by at several these sites, we studied functional consequences removal Thr669 Ser671 using site-directed mutagenesis. mutant expressed in mouse B82 cells displayed normal binding vivo autophosphorylation was fully active signal transduction as measured EGF-stimulated gene transcription. However, EGF-dependent...