Expression of blood clotting Factor VIII:C gene in capillary endothelial cells

FEBS Letters

Oliveira, Caroline M.¹  Tavárez, José J.¹  Banerjee, Dipak K.¹ 

[1] Department of Biochemistry and Nutrition, School of Medicine, University of Puerto Rico, San Juan, PR 00936-5067 USA

关键词: Blood coagulation factor VIII:C;    Capilliary endothelial cell;    Gene expression;    N-linked glycoprotein;    Disulphide bridge;    Adrenal medulla;   

DOI  :  10.1016/0014-5793(92)80831-Z

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 [₃₅S]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.

【 授权许可】

Unknown