Processing of fusion proteins with immobilized factor Xa.
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.
Assouline
Z
Z
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
Graham
R
R
Miller
R C
RC
Warren
A J
AJ
Kilburn
D G
DG
D000345:Affinity Labels; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002482:Cellulose / Q000378:metabolism; D002522:Chlorocebus aethiops; D006224:Cricetinae; D015951:Factor Xa / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*
D000345:Affinity Labels; D011993:Recombinant Fusion Proteins; D002482:Cellulose; D015951:Factor Xa