High-sensitivity detection and quantitative analysis of native protein-protein interactions and multiprotein complexes by flow cytometry.

Adam G. Schrum, Diana Gil, Elaine P. Dopfer, David L. Wiest, Laurence A. Turka, Wolfgang W A Schamel, E. Palmer

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Most mechanisms of cell development, physiology, and signal transduction are controlled by protein-protein interactions. Immunoprecipitation of multiprotein complexes detected by flow cytometry (IP-FCM) is a means to quantitatively measure these interactions. The high sensitivity of this method makes it useful even when very little biomaterial is available for analysis, as in the case of rare primary cell subsets or patient samples. Detection of the T cell antigen receptor associated with the CD3 multiprotein complex from as few as 300 primary murine T cells is presented as an example. The method is compatible with quantitative flow cytometry techniques, making it possible to estimate the number of coimmunoprecipitated molecules. Both constitutive and inducible protein-protein interactions can be analyzed, as illustrated in related methodology using glutathione S-transferase-fusion protein pull-down experiments. IP-FCM represents a robust, quantitative, biochemical technique to assess native protein-protein interactions, without requiring genetic engineering or large sample sizes.

Original languageEnglish (US)
JournalScience's STKE : signal transduction knowledge environment
Volume2007
Issue number389
DOIs
StatePublished - Jun 23 2007
Externally publishedYes

Fingerprint

Multiprotein Complexes
Flow cytometry
Flow Cytometry
Chemical analysis
Proteins
CD3 Antigens
Genetic engineering
Cell Physiological Phenomena
Signal transduction
Genetic Engineering
T-cells
Physiology
Biocompatible Materials
T-Cell Antigen Receptor
Glutathione Transferase
Immunoprecipitation
Sample Size
Signal Transduction
Fusion reactions
T-Lymphocytes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

High-sensitivity detection and quantitative analysis of native protein-protein interactions and multiprotein complexes by flow cytometry. / Schrum, Adam G.; Gil, Diana; Dopfer, Elaine P.; Wiest, David L.; Turka, Laurence A.; Schamel, Wolfgang W A; Palmer, E.

In: Science's STKE : signal transduction knowledge environment, Vol. 2007, No. 389, 23.06.2007.

Research output: Contribution to journalArticle

Schrum, Adam G. ; Gil, Diana ; Dopfer, Elaine P. ; Wiest, David L. ; Turka, Laurence A. ; Schamel, Wolfgang W A ; Palmer, E. / High-sensitivity detection and quantitative analysis of native protein-protein interactions and multiprotein complexes by flow cytometry. In: Science's STKE : signal transduction knowledge environment. 2007 ; Vol. 2007, No. 389.
@article{07a698763ce447f7a0af973ac94e3cd1,
title = "High-sensitivity detection and quantitative analysis of native protein-protein interactions and multiprotein complexes by flow cytometry.",
abstract = "Most mechanisms of cell development, physiology, and signal transduction are controlled by protein-protein interactions. Immunoprecipitation of multiprotein complexes detected by flow cytometry (IP-FCM) is a means to quantitatively measure these interactions. The high sensitivity of this method makes it useful even when very little biomaterial is available for analysis, as in the case of rare primary cell subsets or patient samples. Detection of the T cell antigen receptor associated with the CD3 multiprotein complex from as few as 300 primary murine T cells is presented as an example. The method is compatible with quantitative flow cytometry techniques, making it possible to estimate the number of coimmunoprecipitated molecules. Both constitutive and inducible protein-protein interactions can be analyzed, as illustrated in related methodology using glutathione S-transferase-fusion protein pull-down experiments. IP-FCM represents a robust, quantitative, biochemical technique to assess native protein-protein interactions, without requiring genetic engineering or large sample sizes.",
author = "Schrum, {Adam G.} and Diana Gil and Dopfer, {Elaine P.} and Wiest, {David L.} and Turka, {Laurence A.} and Schamel, {Wolfgang W A} and E. Palmer",
year = "2007",
month = "6",
day = "23",
doi = "10.1126/stke.3892007pl2",
language = "English (US)",
volume = "2007",
journal = "Science Signaling",
issn = "1937-9145",
publisher = "American Association for the Advancement of Science",
number = "389",

}

TY - JOUR

T1 - High-sensitivity detection and quantitative analysis of native protein-protein interactions and multiprotein complexes by flow cytometry.

AU - Schrum, Adam G.

AU - Gil, Diana

AU - Dopfer, Elaine P.

AU - Wiest, David L.

AU - Turka, Laurence A.

AU - Schamel, Wolfgang W A

AU - Palmer, E.

PY - 2007/6/23

Y1 - 2007/6/23

N2 - Most mechanisms of cell development, physiology, and signal transduction are controlled by protein-protein interactions. Immunoprecipitation of multiprotein complexes detected by flow cytometry (IP-FCM) is a means to quantitatively measure these interactions. The high sensitivity of this method makes it useful even when very little biomaterial is available for analysis, as in the case of rare primary cell subsets or patient samples. Detection of the T cell antigen receptor associated with the CD3 multiprotein complex from as few as 300 primary murine T cells is presented as an example. The method is compatible with quantitative flow cytometry techniques, making it possible to estimate the number of coimmunoprecipitated molecules. Both constitutive and inducible protein-protein interactions can be analyzed, as illustrated in related methodology using glutathione S-transferase-fusion protein pull-down experiments. IP-FCM represents a robust, quantitative, biochemical technique to assess native protein-protein interactions, without requiring genetic engineering or large sample sizes.

AB - Most mechanisms of cell development, physiology, and signal transduction are controlled by protein-protein interactions. Immunoprecipitation of multiprotein complexes detected by flow cytometry (IP-FCM) is a means to quantitatively measure these interactions. The high sensitivity of this method makes it useful even when very little biomaterial is available for analysis, as in the case of rare primary cell subsets or patient samples. Detection of the T cell antigen receptor associated with the CD3 multiprotein complex from as few as 300 primary murine T cells is presented as an example. The method is compatible with quantitative flow cytometry techniques, making it possible to estimate the number of coimmunoprecipitated molecules. Both constitutive and inducible protein-protein interactions can be analyzed, as illustrated in related methodology using glutathione S-transferase-fusion protein pull-down experiments. IP-FCM represents a robust, quantitative, biochemical technique to assess native protein-protein interactions, without requiring genetic engineering or large sample sizes.

UR - http://www.scopus.com/inward/record.url?scp=34250684995&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34250684995&partnerID=8YFLogxK

U2 - 10.1126/stke.3892007pl2

DO - 10.1126/stke.3892007pl2

M3 - Article

VL - 2007

JO - Science Signaling

JF - Science Signaling

SN - 1937-9145

IS - 389

ER -