用固定化因子Xa处理融合蛋白。

Processing of fusion proteins with immobilized factor Xa.

作者信息

Assouline Z, Graham R, Miller R C, Warren A J, Kilburn D G

机构信息

Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.

出版信息

Biotechnol Prog. 1995 Jan-Feb;11(1):45-9. doi: 10.1021/bp00031a006.

DOI:10.1021/bp00031a006

PMID:7765987

Abstract

Factor Xa, with a cellulose-binding domain (CBD) fused to the C-terminus of the heavy chain (FXa-CBD), is active in solution and when immobilized on cellulose. A second derivative of factor Xa in which a hexahistidine tail is fused to the C-terminus of the heavy chain (FXa-H6) also retains activity when immobilized, in this case on Ni(2+)-NTA agarose. The stabilities and activities of of FXa-CBD and FXa-H6 immobilized on cellulose and Ni(2+)-NTA agarose, respectively, are similar. Immobilized factor Xa derivatives can be used to remove affinity tags from appropriate fusion proteins without contaminating the desired product with factor Xa.

摘要

凝血因子Xa（Factor Xa），其重链C末端融合了一个纤维素结合结构域（CBD）（FXa-CBD），在溶液中以及固定在纤维素上时均具有活性。凝血因子Xa的第二种衍生物，其中六聚组氨酸尾融合到重链的C末端（FXa-H6），在固定时（在这种情况下固定在Ni(2+)-NTA琼脂糖上）也保留活性。分别固定在纤维素和Ni(2+)-NTA琼脂糖上的FXa-CBD和FXa-H6的稳定性和活性相似。固定化的凝血因子Xa衍生物可用于从适当的融合蛋白上去除亲和标签，而不会使所需产物被凝血因子Xa污染。