Skip to main content

Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit.

Publication ,  Journal Article
Connor, JH; Frederick, D; Huang, HB; Yang, J; Helps, NR; Cohen, PT; Nairn, AC; DePaoli-Roach, A; Tatchell, K; Shenolikar, S
Published in: J Biol Chem
June 23, 2000

Inhibitor-1 (I-1) and inhibitor-2 (I-2) selectively inhibit type 1 protein serine/threonine phosphatases (PP1). To define the molecular basis for PP1 inhibition by I-1 and I-2 charged-to-alanine substitutions in the Saccharomyces cerevisiae, PP1 catalytic subunit (GLC7), were analyzed. Two PP1 mutants, E53A/E55A and K165A/E166A/K167A, showed reduced sensitivity to I-2 when compared with wild-type PP1. Both mutants were effectively inhibited by I-1. Two-hybrid analysis and coprecipitation or pull-down assays established that wild-type and mutant PP1 catalytic subunits bound I-2 in an identical manner and suggested a role for the mutated amino acids in enzyme inhibition. Inhibition of wild-type and mutant PP1 enzymes by full-length I-2(1-204), I-2(1-114), and I-2(36-204) indicated that the mutant enzymes were impaired in their interaction with the N-terminal 35 amino acids of I-2. Site-directed mutagenesis of amino acids near the N terminus of I-2 and competition for PP1 binding by a synthetic peptide encompassing an I-2 N-terminal sequence suggested that a PP1 domain composed of amino acids Glu-53, Glu-55, Asp-165, Glu-166, and Lys-167 interacts with the N terminus of I-2. This defined a novel regulatory interaction between I-2 and PP1 that determines I-2 potency and perhaps selectivity as a PP1 inhibitor.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

June 23, 2000

Volume

275

Issue

25

Start / End Page

18670 / 18675

Location

United States

Related Subject Headings

  • Sequence Homology, Amino Acid
  • Saccharomyces cerevisiae
  • Protein Phosphatase 1
  • Protein Binding
  • Phosphoprotein Phosphatases
  • Mutagenesis, Site-Directed
  • Molecular Sequence Data
  • Models, Chemical
  • Enzyme Inhibitors
  • DNA Primers
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Connor, J. H., Frederick, D., Huang, H. B., Yang, J., Helps, N. R., Cohen, P. T., … Shenolikar, S. (2000). Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit. J Biol Chem, 275(25), 18670–18675. https://doi.org/10.1074/jbc.M909312199
Connor, J. H., D. Frederick, H. B. Huang, J. Yang, N. R. Helps, P. T. Cohen, A. C. Nairn, A. DePaoli-Roach, K. Tatchell, and S. Shenolikar. “Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit.J Biol Chem 275, no. 25 (June 23, 2000): 18670–75. https://doi.org/10.1074/jbc.M909312199.
Connor JH, Frederick D, Huang HB, Yang J, Helps NR, Cohen PT, et al. Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit. J Biol Chem. 2000 Jun 23;275(25):18670–5.
Connor, J. H., et al. “Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit.J Biol Chem, vol. 275, no. 25, June 2000, pp. 18670–75. Pubmed, doi:10.1074/jbc.M909312199.
Connor JH, Frederick D, Huang HB, Yang J, Helps NR, Cohen PT, Nairn AC, DePaoli-Roach A, Tatchell K, Shenolikar S. Cellular mechanisms regulating protein phosphatase-1. A key functional interaction between inhibitor-2 and the type 1 protein phosphatase catalytic subunit. J Biol Chem. 2000 Jun 23;275(25):18670–18675.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

June 23, 2000

Volume

275

Issue

25

Start / End Page

18670 / 18675

Location

United States

Related Subject Headings

  • Sequence Homology, Amino Acid
  • Saccharomyces cerevisiae
  • Protein Phosphatase 1
  • Protein Binding
  • Phosphoprotein Phosphatases
  • Mutagenesis, Site-Directed
  • Molecular Sequence Data
  • Models, Chemical
  • Enzyme Inhibitors
  • DNA Primers