Proteome analysis of activated murine B-lymphocytes

Abstract

Proteins extracted from murine B-lymphocytes after in vitro stimulation by lipopolysaccharide were separated by two-dimensional (2-D) polyacrylamide gel electrophoresis and analyzed by matrix assisted laser desorption/ionization mass spectrometry. Structural information on the protein entities from 153 spots was obtained. Since many of these spots occur as members of spot families, a smaller number --98 genes-- was found to be coding for the identified spots. The elucidated proteins belong to groups of functional categories; we found 26 enzymes, 36 regulatory proteins, 15 chaperones, 15 structural proteins, 4 immunoglobulins, 1 ribosomal and 1 histone protein. A comparison between expected and observed molecular masses yields a good correlation for the majority of the compared spot entities. This set of proteins now identified in the context of a lymphocyte 2-D gel pattern should advance further studies on lymphocyte functions.

Proteome analysis of B‐cell maturation

“…We followed the overall changes in protein abundance induced by mIgM engagement and compared the proteomic data to the corresponding DNA-microarray studies. To observe long-term trends, we followed the proteomic profile at eight time points during 5 d. Proteomic analyses of distinct stages of B-cell maturation have been reported both for mice and human cells [28][29][30][31][32]. Our observations of the proteins involved in B-cell maturation are consistent with the previous transcriptomic and proteomic studies and provide time course data to follow the events after BCR stimulation.…”

“…The first 2-DE analyses of differentiating B-cells were performed in the early 1980s, but at that time proteins could not be identified [79,80]. Since the beginning of the 21st century, with the advent of MS-based protein identification, some 2-DE-based proteomic analyses of distinct stages of B-cell maturation have been reported for cultured human [32] and mice cells [28,30,31,81]. Proteomic analysis has been applied also to healthy blood donors in order to determine the proteomic expression of the different T-and B-cell populations [29] and to human lymphoid neoplasm to distinguish histological subtypes of Hodgkin's lymphoma, natural killer cell lymphoma and B and T cell malignancies [82].…”

Comparative proteomic analysis of GS‐NS0 murine myeloma cell lines with varying recombinant monoclonal antibody production rate

“…Some of the most abundant proteins identified in GS-NS0 cells were (1) elongation factor 1a (EF11) that promotes the GTP-dependent binding of aminoacyl-tRNA to the A-site of ribosomes during protein biosynthesis, (2) glycolytic enzymes: glyceraldehyde-3-phosphate dehydrogenase (G3P) and alpha-enolase (ENOA), and (3) cytosolic and ER resident molecular chaperones: immunoglobulin binding protein (BiP) and heat shock cognate 71 kDa protein (HSC7). Previous proteomic studies have shown that cytosolic and ER resident molecular chaperones are among the most highly abundant proteins in CHO cells (Champion et al, 1999) and activated B-cells (Frey et al, 2000;van Anken et al, 2003). The predominant ER lumenal chaperone, BiP, was present as a main isoform (BiPb) with evidence of a subsidiary, more acidic form (BiPa).…”

Functional proteomics: The goalposts are moving

“…Consequently, ERp29 appears generally susceptible to a subtle post-translational modification such as partial deamidation. The broad range of reported M r values, extending 10% above and below Before detailed characterization of purified ERp29 (left panel), several groups had identified ERp29-like protein spots by partial sequence analysis [32,[47][48][49][51][52][53]. Uncertain relationships existed between these proteins (grey circles depict the reported 2-DE coordinates), and with ERp29/ERp28 cDNAs isolated from rat and human (triangles depict coordinates of inferred mature proteins).…”