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Identification of swiprosin 1 in human lymphocytes
Author(s) -
Vuadens Françoise,
Rufer Nathalie,
Kress Annegret,
Corthésy Patricia,
Schneider Philippe,
Tissot JeanDaniel
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300779
Subject(s) - microbiology and biotechnology , blot , biology , gel electrophoresis , tandem mass spectrometry , cd8 , cd19 , chemistry , mass spectrometry , silver stain , gene , antigen , biochemistry , chromatography , flow cytometry , genetics
Abstract The aim of this work was to identify a new protein that discriminated CD8 from CD4 and CD19 lymphocytes. Proteins were separated by high‐resolution two‐dimensional electrophoresis. After silver staining, the gel images were captured with a laser densitometer, and studied with a dedicated software. This study confirmed the presence of two spots that appeared to be preferentially associated with CD8 lymphocytes, and mass spectrometry analyzes (liquid chromatography‐tandem mass spectrometry, LC‐MS/MS) identified six peptides for one spot and four for the other. The peptide sequences corresponded to an unknown protein that we named swiprosin 1 (Swiss‐Prot Q96C19). Molecular analysis (reverse transcriptase‐polymerase chain reaction, RT‐PCR) and Northern blots confirmed that the gene expression was increased in purified populations of CD8 lymphocytes, when compared to CD19 and CD4 lymphocytes. Database mining revealed that swiprosin 1 contains two potential EF‐hand domains, and therefore may have a role in calcium signaling. Its predominant presence in CD8 lymphocytes suggests that it may be involved in functions that are important for cytotoxic lymphocytes.