A5026986944- Advanced Proteomics Techniques and Applications
- Cholesterol and Lipid Metabolism
- Peroxisome Proliferator-Activated Receptors
- Cellular transport and secretion
- Mass Spectrometry Techniques and Applications
- Genomics and Phylogenetic Studies
- RNA and protein synthesis mechanisms
- Enzyme Structure and Function
- Ubiquitin and proteasome pathways
- Glycosylation and Glycoproteins Research
- Drug Transport and Resistance Mechanisms
- Endoplasmic Reticulum Stress and Disease
- Cardiac Ischemia and Reperfusion
- Inflammatory Biomarkers in Disease Prognosis
- Antiplatelet Therapy and Cardiovascular Diseases
- Alzheimer's disease research and treatments
- Inflammasome and immune disorders
- Sphingolipid Metabolism and Signaling
- Mitochondrial Function and Pathology
- Trace Elements in Health
- Machine Learning in Bioinformatics
- Cardiac and Coronary Surgery Techniques
- Bacterial Genetics and Biotechnology
- Lipid metabolism and disorders
- Studies on Chitinases and Chitosanases
Concord Repatriation General Hospital
2014-2021
Anzac Research Institute
2014-2020
The University of Sydney
2014-2020
Macquarie University
1998-2014
UNSW Sydney
2006-2009
University of Verona
2003
Proteome Sciences (United Kingdom)
2001
Australian Genome Research Facility
1998
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 Oxidative stress and mitochondrial dysfunction have been linked to neurodegenerative disorders such as Parkinson's Alzheimer's disease. However, it is not yet understood how endogenous oxidative may result in dysfunction. Most prior studies tested paradigms mitochondria through either chemical inhibition of specific components the respiratory chain, or adding an exogenous insult hydrogen peroxide paraquat directly damage mitochondria. In contrast, mice that lack superoxide dismutase...
Abundant and hydrophilic nonmembrane proteins with isoelectric points below pH 8 are the predominant identified in most proteomics projects. In yeast, however, low-abundance make up 80% of predicted proteome, approximately 50% have pI's above 30% yeast ORFs to encode membrane at least 1 trans-membrane span. By applying highly solubilizing reagents fractionation a fraction we purified 780 protein isoforms, representing 323 gene products, including 28% low abundance 49% or associated proteins....
Abstract Background Arsenic and cadmium are widely distributed in nature pose serious threats to the environment human health. Exposure these nonessential toxic metals may result a variety of diseases including cancer. However, arsenic toxicity targets cellular systems contributing tolerance acquisition not fully known. Results To gain insight into metal action mechanisms, we carried out genome-wide screening Saccharomyces cerevisiae haploid homozygous diploid deletion mutant collections...
Abstract The development of automated, high throughput technologies for the rapid identification proteins is essential large‐scale proteome projects. While a degree automation already exists in some stages protein process, such as automated acquisition matrix assisted laser desorption ionisation — time flight (MALDI‐TOF) mass spectra, efficient interfaces between different are still lacking. We report highly integrated system large scale separated by two‐dimensional gel electrophoresis...
Cholesterol-loaded foam cell macrophages are prominent in atherosclerotic lesions and play complex roles both inflammatory signaling lipid metabolism, which underpinned by large scale reprogramming of gene expression. We performed a microarray study primary human that showed transcription the sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is up-regulated after cholesterol loading. SMPDL3A protein expression secretion from stimulated loading, liver X receptor ligands, cyclic AMP,...
Amyloid-β (Aβ)-containing plaques are a major neuropathological feature of Alzheimer's disease (AD). The two isoforms Aβ peptide associated with AD Aβ40 and Aβ42, which the latter is highly prone to aggregation. Increased presence aggregation intracellular Aβ42 peptides an early event in progression. Improved understanding cellular processes affecting may have implications for development therapeutic strategies. fused green fluorescent protein (Aβ42-GFP) was expressed ∼4600 mutants...
Abstract Colchicine inhibits coronary and cerebrovascular events in patients with artery disease (CAD), although known to have anti-inflammatory properties, its mechanisms of action are incompletely understood. In this study, we investigated the effects colchicine on platelet activation a particular focus via collagen glycoprotein (GP)VI receptor, P2Y 12 procoagulant formation. Therapeutic concentrations vitro (equivalent plasma levels) significantly decreased aggregation whole blood rich...
Apolipoprotein α-I (apoA-I) is themajor component of HDLandcentral to the ability HDLto stimulate ATP-binding cassette transporter A1 (ABCA1)-dependent, antiatherogenic export cholesterol from macrophage foam cells, a key player in pathology atherosclerosis. Cell-mediated modifications apoA-I, such as chlorination, nitration, oxidation, and proteolysis, can impair its function, although it unknown whether macrophages themselves contribute modifications. To investigate this, human...
Cyclosporin A (CsA) is an immunosuppressant that inhibits protein phosphatase 2B (PP2B/calcineurin) and associated with hyperlipidemia, decreased cholesterol efflux via ATP-binding cassette transporter A1 (ABCA1), increased risk of atherosclerosis. Apolipoprotein E (apoE) important regulator lipid metabolism atherosclerosis, the secretion which from human macrophages regulated by serine/threonine kinase (PKA) intracellular calcium (Ca2+) (Kockx, M., Guo, D. L., Huby, T., Lesnik, P., Kay, J.,...
Expressed Sequence Tags (ESTs) are an invaluable resource for protein identification and characterisation in proteomics. They allow proteins to be identified the absence of genome sequence data. When EST sequences used identification, they usually first processed into contigs reduce redundancy generate longer from overlapping ESTs. However, process generating may accidentally group biologically meaningful isoforms together. Here we report means discovering how use this information framework...
Abstract 2‐DE separations of protein extracts sometimes have problems with poor resolution and streaking. This problem is particularly apparent microorganisms, most notably those a large cell wall. Here we describe novel, rapid protocol for the extraction microorganisms in acidic conditions, leading to increased 2‐D gel quality. The efficiency demonstrated bacteria, Escherichia coli Bacillus subtilis ; fungus, Trichoderma harzianum yeast, Saccharomyces cerevisiae . We also demonstrate using...
Secretion of spore coat proteins from the prespore secretory vesicles (PSVs) in Dictyostelium discoideum is a signal mediated event that underlies terminal cell differentiation, and represents an important case developmentally regulated secretion. In order to study biochemical mechanisms govern fusion PSVs with plasma membrane subsequent secretion their cargo, we purified this organelle cells. Analysis protein extracts highly indicated that, addition cargo structural proteins, many more are...
Dynamins are fission proteins that mediate endocytic and exocytic membrane events pharmacological therapeutic targets. These studies investigate whether dynamin II regulates constitutive protein secretion show for the first time inhibition of decreases apolipoprotein E (apoE) several other constitutively secreted from primary human macrophages. Inhibitors target recruitment to membranes (MiTMABs) or directly GTPase domain (Dyngo Dynole series), dose- time- dependently reduced apoE. SiRNA...
Platelets are critical in mediating both rapid responses to injury and the development progression of coronary disease. Several studies have shown that, after prolonged exposure agonists, they produce release inflammatory mediators including interleukin-1β (IL-1β), via classical pathway (NLRP3 inflammasome caspase-1 cleavage active IL-1β) as described for leukocytes. This study aimed determine whether there is IL-1β response soluble platelet agonists such NLRP3- caspase-1-dependent. Using...
Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which secreted N-linked glycoprotein. SMPDL3A highly homologous to acid sphingomyelinase (aSMase), but unlike aSMase cannot cleave sphingomyelin. Rather, hydrolyzes nucleotide tri- and diphosphates their derivatives. While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of biology, may give insight into its function in vivo, remain...