Lentil lectin enriched microsomes from the plasma membrane of the human B-lymphocyte cell line H2LCL carry a heavy load of type-1 porin

Biol Chem. 1998 Dec;379(12):1419-26. doi: 10.1515/bchm.1998.379.12.1419.

Abstract

Using an established biochemical approach, five subcellular fractions of human B lymphocytes were prepared by differential centrifugation. Crude membranes were passed over a lentil lectin column to enrich carbohydrate-coated cell surface microsomes. The lectin-bound fraction contained a high amount of plasma membrane-derived microsomes as indicated by cell surface markers. All subcellular fractions in Western blots proved to contain distinct but variable amounts of porin. There was a strong increase in porin content from crude membranes to plasma membrane-derived vesicles. The porin content of this fraction appeared to be higher than that of mitochondria. In the final step the plasma membrane-derived microsome fraction proved to be devoid of contamination by outer mitochondrial membranes, as revealed by antibodies against the established markers MAO B and Tom20 applied in Western blots. These data prove the extramitochondrial expression of human type-1 porin/ type-1 VDAC.

MeSH terms

  • B-Lymphocytes / metabolism*
  • Blotting, Western
  • Cell Compartmentation
  • Cell Line, Transformed
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Lectins / administration & dosage*
  • Microsomes / metabolism*
  • Plant Lectins*
  • Porins / metabolism*

Substances

  • Lectins
  • Plant Lectins
  • Porins
  • lentil lectin