Histidyl phosphorylation and dephosphorylation of P36 in rat liver extract

Kiyoto Motojima, S. Goto

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Protein histidine kinase (Motojima, K., and Goto, S. (1993) FEBS Lett. 319, 75-79) and phosphatase in rat liver extract were characterized. The histidine kinase was recovered mostly in the membrane and the phosphatase in the soluble fraction. The kinase and its substrate 36-kDa protein (P36) were co-solubilized from the membrane under conditions in which most of the other kinases, and their substrate proteins were not solubilized. The solubilized kinase and P36 were co-eluted after high pressure liquid chromatography gel filtration, showing an apparent molecular mass of 70-75 kDa. They were also co-eluted after ion exchange chromatography. These characteristics, together with its complete resistance to genistein, indicate that the rat liver histidine kinase is not cognate to the yeast enzyme (Huang, J., Nasr, M., Kim, Y., and Matthews, H. R. (1992) J. Biol. Chem. 267, 15511-15515). The phosphatase that dephosphorylates histidyl-phosphorylated P36 was also studied using rat liver subcellular fractions and in vitro phosphorylated P36 as the substrate. The characteristics of the phosphatase, that is, 1) Mg2+ requirement for activity, 2) apparent molecular mass of 45 kDa by high performance liquid chromatography gel filtration, and 3) resistance to 100 μM okadaic acid, suggest that the primary phosphatase active in vitro is protein phosphatase 2C.

Original languageEnglish
Pages (from-to)9030-9037
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number12
Publication statusPublished - Mar 25 1994

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Liver Extracts
Phosphorylation
Phosphoric Monoester Hydrolases
Rats
Phosphotransferases
Histidine
Molecular mass
Gel Chromatography
Liver
High Pressure Liquid Chromatography
Substrates
Okadaic Acid
Gels
High pressure liquid chromatography
Subcellular Fractions
Membranes
Genistein
Ion Exchange Chromatography
Proteins
Phosphoprotein Phosphatases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Histidyl phosphorylation and dephosphorylation of P36 in rat liver extract. / Motojima, Kiyoto; Goto, S.

In: Journal of Biological Chemistry, Vol. 269, No. 12, 25.03.1994, p. 9030-9037.

Research output: Contribution to journalArticle

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