Proteomic and biochemical methods to study the cytoskeletome

Richard L. Klemke, Xinning Jiang, Sunkyu Choi, Jonathan A. Kelber

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

The cytoskeleton is fundamental to many cellular functions including cell proliferation, differentiation, adhesion, and migration. It is composed of actin, microtubules, intermediate filaments, and integrin cell surface receptors, which form focal adhesions with the extracellular matrix. These elements are highly integrated in the cell providing a rigid network of interconnected cables and protein scaffolds, which generate force and mechanical support to maintain cell shape and movement. However, the cytoskeleton is not just a simple compilation of static filaments that dictate cell adhesion and morphology - it is highly plastic with the inherent ability to assemble and disassemble in response to diverse and complex cellular cues. Thus, biochemical and proteomic methods are needed to better understand the cytoskeleton network and its dynamic signal transduction functions in health and disease. This chapter describes methods for the biochemical enrichment and mass spectrometry-based proteomic analyses of the cytoskeletome. We also detail how these methods can be used to investigate the cytoskeletome of migrating cells and their purified pseudopodia membrane projections.

Original languageEnglish
Title of host publicationAdhesion Protein Protocols
PublisherHumana Press Inc.
Pages203-218
Number of pages16
ISBN (Print)9781627035378
DOIs
Publication statusPublished - 1 Jan 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1046
ISSN (Print)1064-3745

Fingerprint

Cytoskeleton
Proteomics
Pseudopodia
Focal Adhesions
Aptitude
Intermediate Filaments
Cell Shape
Cell Surface Receptors
Cell Adhesion
Integrins
Microtubules
Plastics
Cell Movement
Cues
Extracellular Matrix
Actins
Cell Differentiation
Signal Transduction
Mass Spectrometry
Cell Proliferation

Keywords

  • Cell migration
  • Cytoskeletome
  • Proteomics
  • Pseudopodium dynamics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Klemke, R. L., Jiang, X., Choi, S., & Kelber, J. A. (2013). Proteomic and biochemical methods to study the cytoskeletome. In Adhesion Protein Protocols (pp. 203-218). (Methods in Molecular Biology; Vol. 1046). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-538-5_12

Proteomic and biochemical methods to study the cytoskeletome. / Klemke, Richard L.; Jiang, Xinning; Choi, Sunkyu; Kelber, Jonathan A.

Adhesion Protein Protocols. Humana Press Inc., 2013. p. 203-218 (Methods in Molecular Biology; Vol. 1046).

Research output: Chapter in Book/Report/Conference proceedingChapter

Klemke, RL, Jiang, X, Choi, S & Kelber, JA 2013, Proteomic and biochemical methods to study the cytoskeletome. in Adhesion Protein Protocols. Methods in Molecular Biology, vol. 1046, Humana Press Inc., pp. 203-218. https://doi.org/10.1007/978-1-62703-538-5_12
Klemke RL, Jiang X, Choi S, Kelber JA. Proteomic and biochemical methods to study the cytoskeletome. In Adhesion Protein Protocols. Humana Press Inc. 2013. p. 203-218. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-538-5_12
Klemke, Richard L. ; Jiang, Xinning ; Choi, Sunkyu ; Kelber, Jonathan A. / Proteomic and biochemical methods to study the cytoskeletome. Adhesion Protein Protocols. Humana Press Inc., 2013. pp. 203-218 (Methods in Molecular Biology).
@inbook{f3561c0f4cd94aed915c459278a725cf,
title = "Proteomic and biochemical methods to study the cytoskeletome",
abstract = "The cytoskeleton is fundamental to many cellular functions including cell proliferation, differentiation, adhesion, and migration. It is composed of actin, microtubules, intermediate filaments, and integrin cell surface receptors, which form focal adhesions with the extracellular matrix. These elements are highly integrated in the cell providing a rigid network of interconnected cables and protein scaffolds, which generate force and mechanical support to maintain cell shape and movement. However, the cytoskeleton is not just a simple compilation of static filaments that dictate cell adhesion and morphology - it is highly plastic with the inherent ability to assemble and disassemble in response to diverse and complex cellular cues. Thus, biochemical and proteomic methods are needed to better understand the cytoskeleton network and its dynamic signal transduction functions in health and disease. This chapter describes methods for the biochemical enrichment and mass spectrometry-based proteomic analyses of the cytoskeletome. We also detail how these methods can be used to investigate the cytoskeletome of migrating cells and their purified pseudopodia membrane projections.",
keywords = "Cell migration, Cytoskeletome, Proteomics, Pseudopodium dynamics",
author = "Klemke, {Richard L.} and Xinning Jiang and Sunkyu Choi and Kelber, {Jonathan A.}",
year = "2013",
month = "1",
day = "1",
doi = "10.1007/978-1-62703-538-5_12",
language = "English",
isbn = "9781627035378",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "203--218",
booktitle = "Adhesion Protein Protocols",

}

TY - CHAP

T1 - Proteomic and biochemical methods to study the cytoskeletome

AU - Klemke, Richard L.

AU - Jiang, Xinning

AU - Choi, Sunkyu

AU - Kelber, Jonathan A.

PY - 2013/1/1

Y1 - 2013/1/1

N2 - The cytoskeleton is fundamental to many cellular functions including cell proliferation, differentiation, adhesion, and migration. It is composed of actin, microtubules, intermediate filaments, and integrin cell surface receptors, which form focal adhesions with the extracellular matrix. These elements are highly integrated in the cell providing a rigid network of interconnected cables and protein scaffolds, which generate force and mechanical support to maintain cell shape and movement. However, the cytoskeleton is not just a simple compilation of static filaments that dictate cell adhesion and morphology - it is highly plastic with the inherent ability to assemble and disassemble in response to diverse and complex cellular cues. Thus, biochemical and proteomic methods are needed to better understand the cytoskeleton network and its dynamic signal transduction functions in health and disease. This chapter describes methods for the biochemical enrichment and mass spectrometry-based proteomic analyses of the cytoskeletome. We also detail how these methods can be used to investigate the cytoskeletome of migrating cells and their purified pseudopodia membrane projections.

AB - The cytoskeleton is fundamental to many cellular functions including cell proliferation, differentiation, adhesion, and migration. It is composed of actin, microtubules, intermediate filaments, and integrin cell surface receptors, which form focal adhesions with the extracellular matrix. These elements are highly integrated in the cell providing a rigid network of interconnected cables and protein scaffolds, which generate force and mechanical support to maintain cell shape and movement. However, the cytoskeleton is not just a simple compilation of static filaments that dictate cell adhesion and morphology - it is highly plastic with the inherent ability to assemble and disassemble in response to diverse and complex cellular cues. Thus, biochemical and proteomic methods are needed to better understand the cytoskeleton network and its dynamic signal transduction functions in health and disease. This chapter describes methods for the biochemical enrichment and mass spectrometry-based proteomic analyses of the cytoskeletome. We also detail how these methods can be used to investigate the cytoskeletome of migrating cells and their purified pseudopodia membrane projections.

KW - Cell migration

KW - Cytoskeletome

KW - Proteomics

KW - Pseudopodium dynamics

UR - http://www.scopus.com/inward/record.url?scp=84934442518&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84934442518&partnerID=8YFLogxK

U2 - 10.1007/978-1-62703-538-5_12

DO - 10.1007/978-1-62703-538-5_12

M3 - Chapter

SN - 9781627035378

T3 - Methods in Molecular Biology

SP - 203

EP - 218

BT - Adhesion Protein Protocols

PB - Humana Press Inc.

ER -