High resolution gel-based proteomics for the analysis of molecular mechanisms
Jens R. Coorssen University of Western Sydney,Australia
| When |
09 Sep, 2011
from
11:00 am to 12:00 pm |
|---|---|
| Where | Auditorium |
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ITQB Seminar
Title: High resolution gel-based proteomics for the analysis of molecular mechanisms
Speaker: Jens R. Coorssen
Affiliation: Foundation Chair of Molecular Physiology
School of Medicine
Head, UWS Molecular Medicine Research Group
Host: Carla Pinheiro-Assistant Researcher at Plant Biochemistry I Laboratory
Abstract:
Two-dimensional gel electrophoresis (2DE) remains widely regarded as a gold-standard for proteomic analyses. Nonetheless, issues with the method have been routinely noted in the ‘review’ literature, although there has been little to substantiate these claims and many do not seem plausible when the 2DE technique is considered from the perspective of its underlying chemistry and that of proteins. As (or perhaps because) gel-based proteomics is a ‘mature’ technology, factors contributing to possible reductions in performance are known and thus it is possible to better optimize ongoing analyses by targeted refinement of the technique. I will review my group’s efforts to quantitatively improve every stage of 2DE analysis, from sample preparation and protein extraction, to in-gel spot ‘fractionation’ to further improve overall protein resolution, through to improved in-gel protein detection approaches for the enhancement of total proteome coverage. As the overall objective of this work is to provide optimal analyses of (patho)physiological mechanisms, I will also provide a brief overview of our applications of this refined 2DE protocol in initial proteomic investigations of human preterm labour, spinal cord injury, and a number of other samples relevant to both basic and clinical sciences. Thus, 2DE remains a rigorous, high-resolution technique for large-scale proteomic analyses, including the dissection of molecular mechanisms. Nevertheless, considering the complexity of native proteomes, it must be remembered that there is no panacea, only pros and cons in all experimental methods. Thus critical, quantitative methodological evaluation and re-evaluation will always lie at the core of the most effective proteomic analyses.
