DMCA
Proteomic-Based Identification of CD4-Interacting Proteins in Human Primary Macrophages
BibTeX
@MISC{André_proteomic-basedidentification,
author = {Rui André and Saraiva Raposo and Benjamin Thomas and Gabriela Ridlova and William James},
title = {Proteomic-Based Identification of CD4-Interacting Proteins in Human Primary Macrophages},
year = {}
}
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Abstract
Background: Human macrophages (Mw) express low levels of CD4 glycoprotein, which is constitutively recycled, and 40– 50 % of its localization is intracellular at steady-state. Although CD4-interacting proteins in lymphoid cells are well characterised, little is known about the CD4 protein interaction-network in human Mw, which notably lack LCK, a Src family protein tyrosine kinase believed to stabilise CD4 at the surface of T cells. As CD4 is the main cellular receptor used by HIV-1, knowledge of its molecular interactions is important for the understanding of viral infection strategies. Methodology/Principal Findings: We performed large-scale anti-CD4 immunoprecipitations in human primary Mw followed by high-resolution mass spectrometry analysis to elucidate the protein interaction-network involved in induced CD4 internalization and degradation. Proteomic analysis of CD4 co-immunoisolates in resting Mw showed CD4 association with a range of proteins found in the cellular cortex, membrane rafts and components of clathrin-adaptor proteins, whereas in induced internalization and degradation CD4 is associated with components of specific signal transduction, transport and the proteasome. Conclusions/Significance: This is the first time that the anti-CD4 co-immunoprecipitation sub-proteome has been analysed in human primary Mw. Our data have identified important Mw cell surface CD4-interacting proteins, as well as regulatory proteins involved in internalization and degradation. The data give valuable insights into the molecular pathways involved
Keyphrases
cd4-interacting protein proteomic-based identification human primary macrophage human primary mw proteomic analysis degradation cd4 methodology principal finding protein interaction-network human mw conclusion significance large-scale anti-cd4 immunoprecipitations low level cd4 co-immunoisolates regulatory protein cd4 protein interaction-network lymphoid cell valuable insight molecular pathway anti-cd4 co-immunoprecipitation sub-proteome induced cd4 internalization main cellular receptor human macrophage cellular cortex clathrin-adaptor protein molecular interaction specific signal transduction membrane raft high-resolution mass spectrometry analysis src family protein tyrosine kinase viral infection strategy first time cd4 glycoprotein induced internalization cd4 association
