Dephosphorylation of distinct sites in myosin light chain by two types of phosphatase in aortic smooth muscle

F. Erdődi, Anikó Rokolya, Michael Bárány, Kate Bárány

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Two types of myosin light chain phosphatase from aortic smooth muscle extract were separated by chromatography on heparin-agarose. The phosphatase which appeared in the flow-through fractions had low activity on actomyosin, its apparent molecular mass was 260 kDa and upon ethanol treatment it generated a catalytic subunit with an apparent molecular mass of 36-39 kDa as determined by gel filtration. This phosphatase preferentially dephosphorylated the α-subunit of phosphorylase kinase and its phosphorylase phosphatase activity was not inhibited by heparin, inhibitor-1 or inhibitor-2. The phosphatase retained by heparin-agarose had high activity on actomyosin, its apparent molecular mass was 150 kDa and upon ethanol treatment it generated a catalytic subunit with an apparent molecular mass of 39-42 kDa. It preferentially dephosphorylated the β-subunit of phosphorylase kinase and its phosphorylase phosphatase activity was not inhibited by heparin, inhibitor-1 or inhibitor-2. Myosin light chain was phosphorylated by myosin light chain kinase in peptides AB (Ser-P) and CD (Thr-P), and/or by protein kinase C in peptides E (Ser-P) and F (Thr.P) as determined by one-dimensional phosphopeptide mapping. The catalytic subunit of heparin-agarose flow-through phosphatase preferentially dephosphorylated peptide F over peptides AB, CD and E in both isolated light chain and actomyosin. The catalytic subunit of heparin-agarose bound phosphatase could effectively dephosphorylate all sites in isolated light chain, whereas it was less effective on dephosphorylation of peptide E in actomyosin.

Original languageEnglish
Pages (from-to)67-74
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1011
Issue number1
DOIs
Publication statusPublished - Mar 28 1989

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Myosin-Light-Chain Phosphatase
Actomyosin
Phosphoric Monoester Hydrolases
Smooth Muscle
Catalytic Domain
Phosphorylase Phosphatase
Phosphorylase Kinase
Heparin
Ethanol
Myosin-Light-Chain Kinase
Light
Phosphopeptides
Myosin Light Chains
Protein Kinase C
Gel Chromatography
Chromatography
Peptides
heparin-sepharose

Keywords

  • (Aortic smooth muscle)
  • Multiple phosphorylation
  • Myosin light chain
  • Myosin light chain phosphatase

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Dephosphorylation of distinct sites in myosin light chain by two types of phosphatase in aortic smooth muscle. / Erdődi, F.; Rokolya, Anikó; Bárány, Michael; Bárány, Kate.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1011, No. 1, 28.03.1989, p. 67-74.

Research output: Contribution to journalArticle

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AB - Two types of myosin light chain phosphatase from aortic smooth muscle extract were separated by chromatography on heparin-agarose. The phosphatase which appeared in the flow-through fractions had low activity on actomyosin, its apparent molecular mass was 260 kDa and upon ethanol treatment it generated a catalytic subunit with an apparent molecular mass of 36-39 kDa as determined by gel filtration. This phosphatase preferentially dephosphorylated the α-subunit of phosphorylase kinase and its phosphorylase phosphatase activity was not inhibited by heparin, inhibitor-1 or inhibitor-2. The phosphatase retained by heparin-agarose had high activity on actomyosin, its apparent molecular mass was 150 kDa and upon ethanol treatment it generated a catalytic subunit with an apparent molecular mass of 39-42 kDa. It preferentially dephosphorylated the β-subunit of phosphorylase kinase and its phosphorylase phosphatase activity was not inhibited by heparin, inhibitor-1 or inhibitor-2. Myosin light chain was phosphorylated by myosin light chain kinase in peptides AB (Ser-P) and CD (Thr-P), and/or by protein kinase C in peptides E (Ser-P) and F (Thr.P) as determined by one-dimensional phosphopeptide mapping. The catalytic subunit of heparin-agarose flow-through phosphatase preferentially dephosphorylated peptide F over peptides AB, CD and E in both isolated light chain and actomyosin. The catalytic subunit of heparin-agarose bound phosphatase could effectively dephosphorylate all sites in isolated light chain, whereas it was less effective on dephosphorylation of peptide E in actomyosin.

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