PMID- 3082370
OWN - NLM
STAT- MEDLINE
DCOM- 19860509
LR  - 20190609
IS  - 0006-3002 (Print)
IS  - 0006-3002 (Linking)
VI  - 886
IP  - 1
DP  - 1986 Apr 8
TI  - Characterization of the membrane-bound protein kinase C and its substrate 
      proteins in canine cardiac sarcolemma.
PG  - 152-61
AB  - Cardiac sarcolemma was purified from canine ventricles. Enrichment of the 
      sarcolemmal membranes was demonstrated by the high (Na+ + K+)-ATPase activity of 
      28.0 +/- 1.5 mumol Pi/mg protein per h and the high concentration of muscarinic 
      receptors with the Bmax of 8.2 +/- 2.5 pmol/mg protein as determined by [3H]QNB 
      binding. The purified sarcolemma also contains significant levels of a 
      membrane-bound Ca2+ and phospholipid-dependent protein kinase (protein kinase C). 
      To elucidate the protein kinase C activity in sarcolemma, a prior incubation of 
      the membranes with EGTA and Triton X-100 was necessary. The specific activity of 
      protein kinase C was found to be 131.4 pmol Pi/mg per min, in the presence of 
      6.25 micrograms phosphatidylserine and 0.5 mM CaCl2. Treatment of sarcolemma with 
      12-O-tetradecanoylphorbol 13-acetate (TPA) and phorbol 12,13-dibutyrate (PBu2) 
      resulted in a concentration-dependent activation of protein kinase C activity. 
      The effect of TPA and PBu2 on protein kinase C in sarcolemma was independent of 
      exogenous Ca2+ and phosphatidylserine. Polymyxin B inhibited 
      phorbol-ester-induced activation of protein kinase C activity. The distribution 
      of protein kinase C in the cytosolic fraction was also examined. The specific 
      activity of the kinase in the cytosolic fraction was 59.7 pmol Pi/mg per min. 
      However, the total protein kinase C activity in the cytosol was 213500 pmol 
      Pi/min, compared to that of 1025 pmol Pi/min in the sarcolemma isolated from 
      approx. 100 g of canine ventricular muscle. Several endogenous proteins in 
      cardiac sarcolemma were phosphorylated in the presence of Ca2+ and 
      phosphatidylserine. The major substrates for protein kinase C were proteins of Mr 
      94 000, 87 000, 78 000, 51 000, 46 000, 11 500 and 10 000. Most of these 
      substrate proteins have not been identified before. Other proteins of Mr 38 000, 
      31 000 and 15 000 were markedly phosphorylated in the presence of Ca2+ only. 
      Phosphorylation of phospholamban (Mr 27 000 and 11 000) was also stimulated in 
      the presence of Ca2+ and phosphatidylserine, but the low Mr form of phospholamban 
      was distinct from two other low Mr substrate proteins for protein kinase C. 
      Polymyxin B was more selective in inhibiting the protein kinase C dependent 
      phosphorylation. On the other hand, trifluoperazine selectively inhibited the 
      phosphorylation of phospholamban and Mr 15 000 protein. Although the exact 
      function of this kinase is unknown, based on these observations, we believe that 
      protein kinase C in the cardiac sarcolemma may play an important role in the 
      cell-surface-signal regulated cardiac function.
FAU - Yuan, S
AU  - Yuan S
FAU - Sen, A K
AU  - Sen AK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Biochim Biophys Acta
JT  - Biochimica et biophysica acta
JID - 0217513
RN  - 0 (Detergents)
RN  - 0 (Phosphoproteins)
RN  - 3WJQ0SDW1A (Polyethylene Glycols)
RN  - 526U7A2651 (Egtazic Acid)
RN  - 6581-06-2 (Quinuclidinyl Benzilate)
RN  - 9002-93-1 (Octoxynol)
RN  - EC 2.7.11.13 (Protein Kinase C)
SB  - IM
MH  - Animals
MH  - Cell Fractionation
MH  - Detergents/pharmacology
MH  - Dogs
MH  - Egtazic Acid/pharmacology
MH  - Kinetics
MH  - Molecular Weight
MH  - Myocardium/*enzymology
MH  - Octoxynol
MH  - Phosphoproteins/isolation & purification
MH  - Phosphorylation
MH  - Polyethylene Glycols/pharmacology
MH  - Protein Kinase C/*metabolism
MH  - Quinuclidinyl Benzilate/metabolism
MH  - Sarcolemma/*enzymology/ultrastructure
MH  - Substrate Specificity
EDAT- 1986/04/08 00:00
MHDA- 1986/04/08 00:01
CRDT- 1986/04/08 00:00
PHST- 1986/04/08 00:00 [pubmed]
PHST- 1986/04/08 00:01 [medline]
PHST- 1986/04/08 00:00 [entrez]
AID - 0167-4889(86)90221-1 [pii]
AID - 10.1016/0167-4889(86)90221-1 [doi]
PST - ppublish
SO  - Biochim Biophys Acta. 1986 Apr 8;886(1):152-61. doi: 
      10.1016/0167-4889(86)90221-1.