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Effect of okadaic acid on protein phosphorylation patterns of chicken myogenic cells with special reference to
W Hemmer1, M Skarli, J C Perriard
1Institute for Cell Biology, ETH-Hönggerberg, Zürich, Switzerland.
Abstract:
Okadaic acid and other agents affecting cellular phosphorylation and dephosphorylation processes profoundly changed the phosphoprotein pattern of 32Pi-labelled chicken embryonic skeletal muscle cells. The phosphorylation states of proteins in the lower molecular weight range were especially increased. Immunoprecipitation of cellular extracts with anti-creatine kinase antibodies enabled us to identify creatine kinase (CK) phosphoproteins. B-CK was phosphorylated after treating the cultures with 1-oleoyl-2-acetyl-sn-glycerol, dibutyryl-cAMP, okadiac acid and combinations thereof, but not with 1,2-dioleoyl-sn-glycerol. M-CK was also shown to be phosphorylated. The results indicated that in vivo, CK isoforms in muscle are subjected to control mediated by phosphorylation and dephosphorylation processes.
Insights
Cellular phosphorylation and dephosphorylation processes significantly alter muscle phosphoprotein patterns. Creatine kinase (CK) isoforms, specifically B-CK and M-CK, are shown to be phosphorylated in vivo, indicating regulatory control.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Phosphorylation and dephosphorylation are critical cellular regulatory mechanisms.
- Skeletal muscle cells possess complex phosphoprotein patterns influenced by various signaling pathways.
Purpose of the Study:
- To investigate the impact of agents affecting phosphorylation/dephosphorylation on chicken embryonic skeletal muscle phosphoproteins.
- To identify specific phosphoproteins, particularly creatine kinase (CK) isoforms, regulated by these processes.
Main Methods:
- Utilized 32Pi labeling to track protein phosphorylation.
- Employed okadaic acid and other agents to modulate cellular phosphorylation states.
- Used immunoprecipitation with anti-creatine kinase antibodies to isolate and identify CK phosphoproteins.
Main Results:
- Okadaic acid and related agents profoundly altered the phosphoprotein pattern in muscle cells.
- Increased phosphorylation was observed in lower molecular weight proteins.
- Both B-CK and M-CK isoforms were identified as phosphoproteins following specific treatments.
Conclusions:
- In vivo, creatine kinase (CK) isoforms in muscle are subject to regulation via phosphorylation and dephosphorylation.
- These findings highlight the dynamic regulation of muscle proteins by cellular signaling pathways.