Studies on human porin: XIII. The type‐1 VDAC ‘Porin 31HL’ biotinylated at the plasmalemma of Trypan blue excluding human B lymphocytes

FEBS Letters

Hellmann, Klaus Peter¹  Hellmann, Thea¹  Reymann, Susanne¹  Thinnes, Friedrich P.¹  Flörke, Harald¹  Götz, Hilde¹  Hilschmann, Norbert¹  Jakob, Christian¹ 

[1] Max-Planck-Institut für experimentelle Medizin, Abt. Immunchemie, Hermann-Rein Str.3, D-37075 Göttingen, Germany

关键词: ‘Porin 31HL’;    VDAC;    Chloride-channels;    Biotin;    Two-dimensional electrophoresis;    VDAC;    voltage-dependent anion-selective channel;    ‘Porin 31 HL’;    31 kDa human lymphocyte porin;    NHS;    N-hydroxysulfosuccinimide-ester of biotin;    NEPHGE;    non-equilibrium pH gradient gel electrophoresis;    SDS-PAGE;    sodium-dodecylsulfate polyacrylamide gel electrophoresis;    FITC;    fluoresceinisothiocyanate;    PMSF;    phenylmethylsulfonylfluoride;    EDTA;    ethylenediaminetetraacetic acid;    PVDF;    polyvinylidendifluoride;    BSA;    bovine serum albumine;    HRP;    horseradish peroxidase;   

DOI  :  10.1016/0014-5793(95)00465-L

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In 1989, we demonstrated for the first time the expression of the VDAC ‘Porin 31HL’ in the plasmalemma of human B lymphocytes, then giving first evidence of a multi-topological expression of VDAC. Meanwhile, data from this and other laboratories support our proposal of a multi-compartment distribution of the channel in mammalian tissues. Here, by biotinylation of plasmalemma-integrated proteins of proven living B lymphocytes, followed by two-dimensional electrophoresis, immuno- and streptavidin affinity blotting, we show that part of the channel molecules can be labelled at the outer membrane of the cells. Thus, by a relevant approach our results invalidate objections concerning putative cross-reactivity of anti-human Type-1 porin antibodies with non-VDAC proteins at the outer cell membrane.

【 授权许可】

Unknown