Molecular cloning, expression, purification and characterization of calcineurin from bovine cardiac muscle.
Calcineurin (CaN), also known as calmodulin-dependent phosphatase, was cloned from bovine cardiac muscle and the deduced amino acid sequences of CaN A revealed that it had an open reading frame of 511 amino acid residues. As compared to bovine brain CaN A, the cardiac enzyme contains a 10 amino acid (ATVEAIEADE) deletion before the autoinhibitory region. A deletion analysis of the catalytic domain revealed a 20% decrease in phosphatase activity when the N-terminal 200 amino acids were removed from CaN A as compared to the wild type enzyme. The C-terminal deletions of CaN A revealed that in addition to the autoinhibitory domain (residues 457-480), additional adjacent residues (407-456) also inhibited CaN activity. These results point to either a second autoinhibitory region within CaN A or an extension of the previously noted autoinhibitory region within the cardiac CaN A enzyme.
Selvakumar
Ponniah
P
Department of Pathology, College of Medicine, University of Saskatchewan, 20 Campus Drive, Saskatoon, Sask., Canada.
Lakshmikuttyamma
Ashakumary
A
Anderson
Deborah H
DH
Sharma
Rajendra K
RK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019703:Calcineurin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D065095:Calcineurin Inhibitors; D002417:Cattle; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000378:metabolism; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion
D065095:Calcineurin Inhibitors; D006657:Histones; D019703:Calcineurin