Nuclear phosphoprotein phosphatase from calf liver

Ming W. Chou, Eng L. Tan, Chung Yang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Calf liver nuclear phosphoprotein phosphate (phosphoprotein phosphohydrolase, EC 3.1.3.16) has been purified approx. 850-fold. The enzyme has a mol. wt. of 34 000 as determined by SDS-polyacrylamide gel electrophoresis. The purified enzyme has a pH optimum between 7.0 and 7.5 with phosphosphorylase, phosphohistones f1 and f2b, and phosphoprotamine as substrates. The enzyme activity towards these substrates follows the order, phosphophosphorylase > phosphohistone f1 > phosphohistone f2b > phopshoprotamine. The Km values toward phosphophosphorylase and phosphohistone f1 are 17 and 28 μM phosphate, respectively. Dephosphorylated histone f1 and orthophosphate are competitive inhibitors of the enzyme with respective Ki values of 11 μM and 4.1. mM. NaCl and divalent metal ions inhibit the enzyme but CaCl2 is slightly stimulatory. It appears that metal ion inhibition occurs at two sites, one on the enzyme and the other on the substrate. The enzyme is also inhibited by NaF and EDTA. Nucleotides bearing the pyrophosphate structure are potent inhibitors of the enzyme while mononucleotides are slightly inhibitory. DNA and other polyions also inhibit the enzyme. The enzyme appears to require free sulfhydryl groups for activity since it is inhibited by N-ethylmaleimide and p-hydroxymercuribenzoate; the latter inhibition can be reversed by mercaptoethanol and dithiothreitol.

Original languageEnglish (US)
Pages (from-to)49-61
Number of pages13
JournalBBA - Enzymology
Volume566
Issue number1
DOIs
StatePublished - Jan 12 1979

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Phosphoprotein Phosphatases
Liver
Enzymes
Phosphates
Enzyme Inhibitors
Metals
Ions
Ethylmaleimide
Mercaptoethanol
Dithiothreitol
Phosphoproteins
Edetic Acid
Histones
Polyacrylamide Gel Electrophoresis
Nucleotides
DNA

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Chou, Ming W. ; Tan, Eng L. ; Yang, Chung. / Nuclear phosphoprotein phosphatase from calf liver. In: BBA - Enzymology. 1979 ; Vol. 566, No. 1. pp. 49-61.
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Nuclear phosphoprotein phosphatase from calf liver. / Chou, Ming W.; Tan, Eng L.; Yang, Chung.

In: BBA - Enzymology, Vol. 566, No. 1, 12.01.1979, p. 49-61.

Research output: Contribution to journalArticle

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