Requirement of protein association with membranes for phosphorylation by protein kinase C.
To clarify the requirement of the association of substrate proteins with phospholipid membranes for phosphorylation by protein kinase C (PKC), we studied the relationship between membrane association of PKC-substrate proteins and their phosphorylation by PKC. In the presence of phosphatidylserine, 12-O-tetradecanoylphorbol-13-acetate induced PKC autophosphorylation in either the presence or the absence of Ca2+, and this phosphorylation was not inhibited by increasing salt concentration (up to 200 mM NaCl). Thus, Ca2+ and ionic strength did not markedly affect the enzymatic activity of PKC. Annexin I required Ca2+ for both its association with phospholipid membranes and phosphorylation by PKC, whereas histone and monomyristilated lysozyme (C14:0-lysozyme) did not. This result indicates that the membrane association of substrates closely correlates with their phosphorylation by PKC. Similar correlation was also observed in the effects of ionic strength on the membrane association of the substrates and their phosphorylation by PKC; increased ionic strength (200 mM NaCl) remarkably inhibited both the membrane association and the phosphorylation of histone and annexin I by PKC but C14:0-lysozyme was not markedly affected. These results suggest that the membrane association of PKC-substrate proteins is a prerequisite for their phosphorylation by PKC. This concept further conforms to the mechanisms of PKC inhibitors; some types of PKC inhibitors are mediated all or in part through inhibition of the substrate-membrane interaction.
Edashige
K
K
Institute for Laboratory Animals, Kochi Medical School, Japan.
Utsumi
T
T
Sato
E F
EF
Ide
A
A
Kasai
M
M
Utsumi
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017302:Annexins; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D009113:Muramidase / Q000378:metabolism; D009994:Osmolar Concentration; D010743:Phospholipids / Q000378:metabolism* / Q000494:pharmacology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
D017302:Annexins; D002135:Calcium-Binding Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D011493:Protein Kinase C; D009113:Muramidase; D002118:Calcium