Publikation: The Lipid Raft Microdomain-Associated Protein Reggie-1/Flotillin-2 is Expressed in Human B Cells and Localized at the Plasma Membrane and Centrosome in PBMCs
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Reggie-1/flotillin-2 is a plasma membrane-associated cytoplasmic protein, which defines non-caveolar raft microdomains. Reggie-1/flotillin-2 is enriched in detergent insoluble (TX100) membrane fractions (DIG), co-localizes with activated GPI-linked proteins and the fyn-kinase in neurons and T cells, and thus apparently participates in the assembly of protein complexes essential for signal transduction. In T cells activated by crosslinking the GPI-linked protein Thy-1 or by crosslinking the ganglioside GM1, reggie-1/flotillin-2 co-localizes with the T cell receptor. To determine whether reggie-1/flotillin-2 is also expressed in B cells, primary B cells from human blood and cell lines representing the developmental stages of pro, pre, mature and plasma B cells were analyzed by Western blotting, RT-PCR and immunofluorescence. Here, we show that reggie-1/flotillin-2 is expressed throughout B cell development, as well as in primary B cells, purified by cell sorting. On non-activated mature B cell Raji cell line we found reggie-1/flotillin-2 are exclusively in the detergent (TX100) insoluble membrane fractions that are staining positive for the raft marker GM1. Immunofluorescence microscopy showed that reggie-1/flotillin-2 is localized at the plasma membrane and marks intracellular spots in PBMCs. Confocal co-localization studies showed that reggie-1/flotillin-2 is associated with the plasma membrane, and the centrosomes (microtubule organizing centers) in these PBMCs. Comparison of reggie-1/flotillin-2 cDNA sequences with the genomic sequence database allowed us to determine the exon/intron structures in mouse and human. The gene organizations are highly conserved suggesting an important function of reggie-1/flotillin-2. Since reggie/flotillin proteins co-cluster with the T cell receptor and fyn kinases upon T cell stimulation, our findings of reggie-1/flotillin-2 in B cells suggest a similar role in B cell function.
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SOLOMON, Samuel, Madhan MASILAMANI, Lawrence RAJENDRAN, Martin BASTMEYER, Claudia STÜRMER, Harald ILLGES, 2002. The Lipid Raft Microdomain-Associated Protein Reggie-1/Flotillin-2 is Expressed in Human B Cells and Localized at the Plasma Membrane and Centrosome in PBMCs. In: Immunobiology. 2002, 205(1), pp. 108-119. ISSN 0171-2985. Available under: doi: 10.1078/0171-2985-00114BibTex
@article{Solomon2002Lipid-7038, year={2002}, doi={10.1078/0171-2985-00114}, title={The Lipid Raft Microdomain-Associated Protein Reggie-1/Flotillin-2 is Expressed in Human B Cells and Localized at the Plasma Membrane and Centrosome in PBMCs}, number={1}, volume={205}, issn={0171-2985}, journal={Immunobiology}, pages={108--119}, author={Solomon, Samuel and Masilamani, Madhan and Rajendran, Lawrence and Bastmeyer, Martin and Stürmer, Claudia and Illges, Harald} }
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