AfCS > AfCS laboratories > Protein Chemistry Laboratory

ALLIANCE FOR CELLULAR SIGNALING

PROTEIN CHEMISTRY LABORATORY

University of Texas Southwestern Medical Center
Dallas, TX

Overview

The Protein Chemistry Laboratory spearheads the proteomics effort of the Alliance for Cellular Signaling (AfCS). The interests of the laboratory center on signaling proteins and changes in covalent modifications. The main focus is currently on phosphoproteins and involves several major efforts.

A primary interest is determining patterns of protein phosphorylation in response to ligand stimulation. The goal is to identify both novel and known phosphoproteins in macrophages. The identified phosphoproteins will be tested for ligand-dependent changes in phosphorylation. The data generated in this effort will expand our knowledge of the signaling molecules involved in ligand responses.

The second focus is to identify the relevant phosphorylation sites on ligand-responsive phosphoproteins. This information will be used to generate phosphospecific antibodies that will then be used to determine quantitative changes in protein phosphorylation.

The Protein Chemistry Laboratory also identifyies proteins isolated in protein-based interaction screens. This information will help expand information on signaling networks by identifying components of signaling complexes.

Major technologies utilized in the Protein Chemistry Laboratory include mass spectrometry, one- and two-dimensional nanoscale liquid chromatography, phosphoprotein enrichment techniques, protein expression, immunoprecipitation, and one- and two-dimensional gel electrophoresis.

Staff

Alliance for Cellular Signaling
Protein Chemistry Laboratory
University of Texas Southwestern Medical Center
5323 Harry Hines Blvd., J5-142
Dallas, Texas 75390-9196
Phone: 214-648-2271
Fax: 214-648-5006

Director	Marc Mumby, Ph.D.
Lead Scientist	Deirdre Brekken, Ph.D.
Lead Scientist	Hongjun Shu, Ph.D.
Research Associate	Farah El Mazouni
Research Assistant	Robert Cox
Research Assistant	Kathleen Lyons
Research Assistant	Deepa Sethuraman
Research Assistant	Laura Draper
Lab Administrator	Kimberly Edwards
Data Manager	Lonnie Sorrells
Lab Aid	Philip Ramirez

Collaborators

The Protein Chemistry Laboratory is collaborating with Cell Signaling Technology (use and development of antibodies): www.cellsignal.com.

Slide Presentation from May 23-26, 2004, AfCS Annual Meeting

• "Protein Phosphorylation: Substrate ID in RAW Cells; SILAC; AQUA Peptides; Bio-Plex", Marc Mumby

Goals for Year 5 (9/04-8/05)

• Validate the SILAC and AQUA peptide methods for quantitative analysis of ligand-stimulated phosphorylation using Erk1 and Akt1 as test cases.
• Produce additional stable populations of RAW 264.7 cells expressing FXM proteins.
• Use the AQUA peptide method in a directed strategy to monitor changes in selected FXM proteins in response to FXM ligands.
• Couple the SILAC method with phosphoprotein and phosphopeptide enrichment methods for global profiling of ligand-induced changes in protein phosphorylation.
• Identify proteins and phosphoproteins as needed by the AfCS.