Methods for pseudopodia purification and proteomic analysis.

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Methods for pseudopodia purification and proteomic analysis. Abstract Text:

Yingchun Wang,Shi-Jian Ding,Wei Wang,Feng Yang,Jon M Jacobs,David Camp,Richard D Smith,Richard L Klemke,Yingchun Wang,Shi-Jian Ding,Wei Wang,Feng Yang,Jon M Jacobs,David Camp,Richard D Smith,Richard L Klemke,

Directional cell migration requires the formation of a dominant pseudopodium in the direction toward which the cell migrates. When a migratory cell is stimulated with a chemoattractant or extracellular matrix (ECM) gradient, it responds with localized amplification of signals on the side facing the gradient. The signals mediate reorganization of the actin-myosin cytoskeleton, leading to morphological polarization of the cell and pseudopodium extension. To identify these signals, we developed an approach to biochemically isolate the pseudopodium from the cell body using 3.0-micrometer porous filters for large-scale quantitative proteomic and phosphoproteomic analysis. Here, we detail the methodology for pseudopodium purification and proteomic analysis. This model system should be widely applicable for the analysis of the pseudopodium proteome from various migratory cell lines, including primary and cancer cell lines stimulated with a diverse array of chemoattractants, ECM proteins, or both.

Methods for pseudopodia purification and proteomic analysis. Publishing Authors By Initials

Y Wang,SJ Ding,W Wang,F Yang,JM Jacobs,D Camp,RD Smith,RL Klemke,Y Wang,SJ Ding,W Wang,F Yang,JM Jacobs,D Camp,RD Smith,RL Klemke,

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Methods for pseudopodia purification and proteomic analysis. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Science's STKE : signal transduction knowledge env

VOLUME: 2007

Page Numbers: pl4

Journal Abbreviation: Sci. STKE

ISSN: 1525-8882

DAY: 21

MONTH: 08

YEAR: 2007

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LANGUAGE: eng

NlmUniqueID: 100964423

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AFFILIATION: Department of Pathology and Moores Cancer Center, University of California, San Diego, La Jolla, CA 92093, USA.

Country: United States

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MEDLINETA: Sci STKE

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