Phosphorylation of myosin light chain from adrenomedullary chromaffin cells in culture.
The myosin-light-chain (MLC) phosphorylation accompanying catecholamine release in chromaffin cells was investigated with the objective of assessing the possible role of this contractile protein in catecholamine secretion. The electrophoretic characteristics of adrenomedullary MLC were determined by immunochemical techniques using two different specific antibodies. The identified 22 kDa phosphoprotein was mainly present in the cytosol, as demonstrated by ultracentrifugation and immunocytochemical analysis. A part of this protein was located on, or close to, the plasma membrane. Cell stimulation by secretagogues resulted in a Ca2(+)-dependent 32P incorporation into MLC, the time course of this process being related to catecholamine release. These findings were supported by a two-dimensional gel-electrophoretic analysis by which means this protein was resolved into two acidic forms. A role for Ca2(+)-calmodulin and Ca2(+)-phospholipid kinases in adrenomedullary MLC phosphorylation is reported. The results obtained suggest a regulatory role for such a protein in the underlying exocytotic event.
Gutierrez
L M
LM
Departamento de Neuroquímica, Facultad de Medicina, Universidad de Alicante, Spain.
Hidalgo
M J
MJ
Palmero
M
M
Ballesta
J J
JJ
Reig
J A
JA
Garcia
A G
AG
Viniegra
S
S
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000313:Adrenal Medulla / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007700:Kinetics; D008970:Molecular Weight; D015879:Myosin Subfragments / Q000302:isolation & purification / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D011188:Potassium / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014268:Trifluoperazine / Q000494:pharmacology
D015879:Myosin Subfragments; D010710:Phosphates; D014268:Trifluoperazine; D000001:Calcimycin; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D013755:Tetradecanoylphorbol Acetate; D011188:Potassium