Recombinant creatine kinase proteins and proposed standards for creatine kinase isoenzyme and subform assays.
Publication/Presentation Date
8-1-1993
Abstract
We developed standards for creatine kinase (CK; EC 2.7.3.2) assays by expressing human CK cDNAs in COS cells. Cells were transiently transfected with full-length cDNAs for CK subunits M and B, individually and in combination; and subsequently, high concentrations of CK activity were present in the cell lysate (1.2 U/mg protein). These proteins exhibited the characteristic isoenzyme-specific electrophoretic mobilities for CK MM and BB isoenzymes. We also produced subforms of CK MM and MB, identical to the modified CK variants produced in plasma after muscle or myocardial injury, by mutating the cDNA for the CK M subunit to delete the carboxy-terminal lysine residue. When this construct was cotransfected with the normal cDNAs for CK M and B, five electrophoretically distinct CK isoenzymes were detected by nondenaturing electrophoresis: MM3, MM2, MM1, MB2, and MB1. These proteins retained 100% of their activity after storage of the cell lysates -20 or 4 degrees C for 3 months.
Volume
39
Issue
8
First Page
1598
Last Page
1601
ISSN
0009-9147
Published In/Presented At
Friedman, D. L., Kesterson, R., Puleo, P., Wu, A. H., & Perryman, M. B. (1993). Recombinant creatine kinase proteins and proposed standards for creatine kinase isoenzyme and subform assays. Clinical chemistry, 39(8), 1598–1601.
Disciplines
Medicine and Health Sciences
PubMedID
8353943
Department(s)
Department of Medicine
Document Type
Article