Ubiquitin-calmodulin conjugating activity from cardiac muscle.
Enzyme activity capable of covalently linking ubiquitin to bovine calmodulin in an ATP-dependent manner has been detected in rabbit cardiac muscle demonstrating that this enzyme occurs not only in reticulocytes but also in other tissues and possibly all tissues and cells which contain calmodulin as intracellular Ca2+-acceptor protein. This is of special interest since a ubiquitin-dependent proteolytic activity could previously not be detected in cardiac muscle. The name ubiquityl-calmodulin synthetase [uCaM-synthetase, ubiquityl:calmodulin ligase (EC 6.3.?.?)] is therefore suggested for this enzyme. In crude cardiac muscle extracts uCaM-Synthetase displays a specific activity of 93 nUnits/mg in comparison to reticulocyte lysate with 270 nUnits/mg as measured by the fluphenazine-Sepharose affinity adsorbent test (FP-test). Analysis of the ubiquitination product (125I-uCaM) by polyacrylamide electrophoresis in the presence of SDS followed by autoradiography reveals a major double band with molecular masses of 27 and 29 kDa (mono-ubiquitination products) respectively. In addition two novel minor bands (17 and 20 kDa) of smaller molecular mass than the monoubiquitination products were detected. These are probably proteolytic breakdown products of uCaM. A model is suggested for a specific function of this synthetase in the Ca2+-dependent breakdown of calmodulin in vertebrate (eukaryotic) cells.
Jennissen
H P
HP
Institut für Physiologie, Ludwig-Maximilians-Universität München.
Laub
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D002474:Cell-Free System; D007700:Kinetics; D008297:Male; D009206:Myocardium / Q000201:enzymology*; D010453:Peptide Synthases / Q000302:isolation & purification / Q000378:metabolism*; D012156:Reticulocytes / Q000378:metabolism; D013737:Testis / Q000378:metabolism; D044764:Ubiquitin-Activating Enzymes; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000378:metabolism*
D002147:Calmodulin; D014452:Ubiquitins; D000255:Adenosine Triphosphate; D044767:Ubiquitin-Protein Ligases; C059404:ubiquitin-calmodulin ligase; D044764:Ubiquitin-Activating Enzymes; D010453:Peptide Synthases