Summary: Mass spectrometry based proteomics approaches can be used in many ways to complement cell biology methodologies. With current state of the art proteomics technology it is possible to accurately locate a protein to sub-cellular locations on a global scale using proteomics, determine its binding partners and absolute abundance.

In this presentation, Kathryn will discuss technologies developed in her laboratory which allow location, binding partners determination and absolute quantification. The first of these technologies, LOPIT (Localization of Organelle Proteins by Isotope Tagging), is a high-throughput proteomics technique that enables protein localization to multiple organelles in a single experiment. Assignment of proteins to sub-cellular compartments is challenging as many organelles cannot be purified to homogeneity. Membranes from the same organelle however, co-sediment in a density gradient, which in turn means that resident proteins of a specific organelle share similar distribution profiles. Sub-cellular localization can therefore be determined by comparing distributions of unknown proteins to those of known organelle markers. Using LOPIT, organelles are first partially separated by density gradient centrifugation and protein distribution patterns quantified using differential stable isotope labeling. Robust quantitative proteomics methods are then applied to accurately quantify distributions of proteins within density gradients. Kathryn has recently shown that they can map the position of protein complexes within organelles using a similar approach, where the density of protein complexes is mapped to organelle locations defined by LOPIT. Finally, they have utilized a method to define the absolute quantitation of proteins of interest by using multiple reaction monitoring methods coupled with the creation of internal standards from an artificial protein. The above methods will be discussed with reference to various biological systems.


SPEAKER BIOGRAPHY:

Kathryn received her PhD in Biochemistry from the University of Sheffield in 1990. She then continued her scientific career as the manager of the Protein and Nucleic Acid Laboratory core facility at the University of Leicester for 11 years, initially starting with Edman degradation, DNA synthesis and HPLC core services and adding peptide synthesis, DNA sequencing and both MALDI-TOF and LC-MS mass spectrometry facilities during this period and assisting in the creation of a cDNA microarray laboratory. In November 2000, she established the Cambridge Center for Proteomics (CCP) of which she is director at the University of Cambridge. This state-of-the-art center collaborates with a large number of groups in the UK and worldwide. The research focus of her group includes development and assessment of methods in quantitative proteomics, especially using these techniques to accurately determine the location of proteins within sub-cellular structures and their interaction partners. The group also collaborates with transcriptomics, metabolomics facilities and bioinformaticians on large-scale system biology approaches. In October 2008 she was appointed to a personal Readership in Protein Chemistry in the Department of Biochemistry, University of Cambridge.  
 
We look forward to seeing you there!
 
Your BAMS Executive Team

Meeting details

Date:	Thursday	April 1st, 2009
Time:	6:00 pm	Social hour, registration (no-host cocktails)
	7:00 pm	Dinner
	8:00 pm	Presentation
Dinner:	featuring:	Dinner selections are A)  Broiled Salmon Champagne Sauce..   B) Roast New Your Strip Steak.   C) Vegetarian Pasta.
Cost:	$30	BAMS members*
	$40	Non-members
	$60	BAMS membership plus dinner cost
	$20	Students only
	$300	2009 Corporate Sponsors
		Dinner reservations required by 7:00 PM, Friday the 27th of March, 2009

*Note:  BAMS memberships are for the calendar year and should be renewed with the first event you attend in 2009.  Dues ($30.00) may be paid while registering for dinner.  If you are unsure of your Membership status, please email Tom Slyker.

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