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Expression of blood clotting Factor VIII:C gene in capillary endothelial cells
Author(s) -
Banerjee Dipak K.,
Tavárez José J.,
Oliveira Caroline M.
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80831-z
Subject(s) - immunoprecipitation , immunoglobulin light chain , microbiology and biotechnology , chemistry , antibody , clotting factor , methionine , gene , biochemistry , biology , medicine , immunology , amino acid
The essential role of Factor VIII:C (FVIII:C, anti‐hemophilia factor A) as a cofactor for Factor IX a ‐dependent activation of Factor X has been established. In this paper, we describe that capillary endothelial cells from bovine adrenal medulla express active FVIII:C gene. Accumulation of FVIII:C in conditioned media from an 8‐day‐old culture is approximately twice as high as that stored in the cell when immunoprecipitated FVIII:C was analyzed for its ability to convert Factor X to Factor X a . Analysis of [ 35 S]methionine‐labeled and immunoprecipitated FVIII:C from cells or conditioned media on SDS‐PAGE under fully denatured conditions indicated that the newly synthesized FVIII:C consists of heavy chain of M r 200,000 and light chain of M r 46,000. The secreted FVIII:C in the non‐reduced condition however, has a molecular weight of 270,000 which suggests that in native protein, the heavy and light chains are held together by S‐S bonds. Furthermore, susceptibility of the immunoprecipitated FVIII:C to N ‐glycanase digestion establishes that the endothelial cells derived FVIII:C contains asparagine‐linked carbohydrate side chains.