TY - JOUR
T1 - Identification of membrane-bound calcium, calmodulin-dependent protein kinase II in canine heart
AU - Jett, Mary Frances
AU - Schworer, Charles M.
AU - Bass, Martha
AU - Soderling, Thomas R.
N1 - Funding Information:
i Supported in part by NIH Grant AM 17808. ’ Present address: University of Pennsylvania School of Nursing, Philadelphia, PA 19104-9096. ’ To whom correspondence would be addressed. * Abbreviations used: CaM, calmodulin; CaM-kinase, calmodulin-dependent protein kinase; Ca*+/CaM-kinase II, Ca2+/CaM-dependent protein kinase II; MLC, myosin light chains; MLC-kinase, myosin light chain kinase; Hepes, 4-(Z-hydroxyethyl)-l-piperazineethanesulfonic acid; EGTA, ethylene glycol bis(@-aminoethyl ether)-N,N’-tetraacetic acid.
PY - 1987/6
Y1 - 1987/6
N2 - Phospholamban, the putative regulatory proteolipid of the Ca2/Mg2+ ATPase in cardiac sarcoplasmic reticulum, was selectively phosphorylated by a Ca2+/calmodulin (CaM)-dependent protein kinase associated with a cardiac membrane preparation. This kinase also catalyzed the phosphorylation of two exogenous proteins known to be phosphorylated by the multifunctional Ca2+/CaM-dependent protein kinase II Ca2+/CaM-kinase II), i.e., smooth muscle myosin light chains and glycogen synthase a. The latter protein was phosphorylated at sites previously shown to be phosphorylated by the purified multifunctional Ca2+/CaM-kinase II from liver and brain. The membrane-bound kinase did not phosphorylate phosphorylase b or cardiac myosin light chains, although these proteins were phosphorylated by appropriate, specific calmodulin-dependent protein kinases added exogenously. In addition to phospholamban, several other membrane-associated proteins were phosphorylated in a calmodulin-dependent manner. The principal one exhibited a Mr of approximately 56,000, a value similar to that of the major protein (57,000) in a partially purified preparation of Ca2+/CaM-kinase II from the soluble fraction of canine heart that was autophosphorylated in a calmodulin-dependent manner. These data indicate that the membrane-bound, calmodulin-dependent protein kinase that phosphorylates phospholamban in cardiac membranes is not a specific calmodulin-dependent kinase, but resembles the multifunctional Ca2+/CaM-kinase II. Our data indicate that this kinase may be present in both the particulate and soluble fractions of canine heart.
AB - Phospholamban, the putative regulatory proteolipid of the Ca2/Mg2+ ATPase in cardiac sarcoplasmic reticulum, was selectively phosphorylated by a Ca2+/calmodulin (CaM)-dependent protein kinase associated with a cardiac membrane preparation. This kinase also catalyzed the phosphorylation of two exogenous proteins known to be phosphorylated by the multifunctional Ca2+/CaM-dependent protein kinase II Ca2+/CaM-kinase II), i.e., smooth muscle myosin light chains and glycogen synthase a. The latter protein was phosphorylated at sites previously shown to be phosphorylated by the purified multifunctional Ca2+/CaM-kinase II from liver and brain. The membrane-bound kinase did not phosphorylate phosphorylase b or cardiac myosin light chains, although these proteins were phosphorylated by appropriate, specific calmodulin-dependent protein kinases added exogenously. In addition to phospholamban, several other membrane-associated proteins were phosphorylated in a calmodulin-dependent manner. The principal one exhibited a Mr of approximately 56,000, a value similar to that of the major protein (57,000) in a partially purified preparation of Ca2+/CaM-kinase II from the soluble fraction of canine heart that was autophosphorylated in a calmodulin-dependent manner. These data indicate that the membrane-bound, calmodulin-dependent protein kinase that phosphorylates phospholamban in cardiac membranes is not a specific calmodulin-dependent kinase, but resembles the multifunctional Ca2+/CaM-kinase II. Our data indicate that this kinase may be present in both the particulate and soluble fractions of canine heart.
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U2 - 10.1016/0003-9861(87)90403-6
DO - 10.1016/0003-9861(87)90403-6
M3 - Article
C2 - 2954508
AN - SCOPUS:0023359654
SN - 0003-9861
VL - 255
SP - 354
EP - 360
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -