Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 289, Issue 21, Pages 14434-14447Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M113.529578
Keywords
-
Categories
Funding
- Chinese Ministry of Science and Technology 973 Program [2012CB911000, 2013CB910700]
- National Natural Science Foundation of China [31110103914, 31070656, 31000342, 31270794]
- Biotechnology and Biological Sciences Research Council (UK) [BB/J021091, H024085/1]
- Medical Research Council (UK) [G0500707]
- Wellcome Trust [094470/Z/10/Z]
- DePaul University Research Council
- Chinese Academy of Sciences [2010T1S11]
- BBSRC [BB/J021091/1, BB/H024085/1] Funding Source: UKRI
- MRC [G0500707] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/J021091/1, BB/H024085/1] Funding Source: researchfish
- British Heart Foundation [RG/09/003/27122] Funding Source: researchfish
- Medical Research Council [G0500707] Funding Source: researchfish
Ask authors/readers for more resources
In the vertebrate immune system, each B-lymphocyte expresses a surface IgM-class B cell receptor (BCR). When cross-linked by antigen or anti-IgM antibody, the BCR accumulates with other proteins into distinct surface clusters that activate cell signaling, division, or apoptosis. However, the molecular composition of these clusters is not well defined. Here we describe a quantitative assay we call selective proteomic proximity labeling using tyramide (SPPLAT). It allows proteins in the immediate vicinity of a target to be selectively biotinylated, and hence isolated for mass spectrometry analysis. Using the chicken B cell line DT40 as a model, we use SPPLAT to provide the first proteomic analysis of any BCR cluster using proximity labeling. We detect known components of the BCR cluster, including integrins, together with proteins not previously thought to be BCR-associated. In particular, we identify the chicken B-lymphocyte allotypic marker chB6. We show that chB6 moves to within about 30 - 40 nm of the BCR following BCR cross-linking, and we show that cross-linking chB6 activates cell binding to integrin substrates laminin and gelatin. Our work provides new insights into the nature and composition of the BCR cluster, and confirms SPPLAT as a useful research tool in molecular and cellular proteomics.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available