PMID- 10318819
OWN - NLM
STAT- MEDLINE
DCOM- 19990617
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 20
DP  - 1999 May 14
TI  - Molecular determinants of nuclear protein phosphatase-1 regulation by NIPP-1.
PG  - 14053-61
AB  - NIPP-1 is a subunit of the major nuclear protein phosphatase-1 (PP-1) in
      mammalian cells and potently inhibits PP-1 activity in vitro. Using yeast
      two-hybrid and co-sedimentation assays, we mapped a PP-1-binding site and the
      inhibition function to the central one-third domain of NIPP-1. Full-length NIPP-1
      (351 residues) and the central domain, NIPP-1(143-217), were equally potent PP-1 
      inhibitors (IC50 = 0.3 nM). Synthetic peptides spanning the central domain of
      NIPP-1 further narrowed the PP-1 inhibitory function to residues 191-200. A
      second, noninhibitory PP-1-binding site was identified by far-Western assays with
      digoxygenin-conjugated catalytic subunit (PP-1C) and included a consensus RVXF
      motif (residues 200-203) found in many other PP-1-binding proteins. The
      substitutions, V201A and/or F203A, in the RVXF motif, or phosphorylation of
      Ser199 or Ser204, which are established phosphorylation sites for protein kinase 
      A and protein kinase CK2, respectively, prevented PP-1C-binding by
      NIPP-1(191-210) in the far-Western assay. NIPP-1(191-210) competed for PP-1
      inhibition by full-length NIPP-1(1-351), inhibitor-1 and inhibitor-2, and
      dissociated PP-1C from inhibitor-1- and NIPP-1(143-217)-Sepharose but not from
      full-length NIPP-1(1-351)-Sepharose. Together, these data identified some of the 
      key elements in the central domain of NIPP-1 that regulate PP-1 activity and
      suggested that the flanking sequences stabilize the association of NIPP-1 with
      PP-1C.
FAU - Beullens, M
AU  - Beullens M
AD  - Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000
      Leuven, Belgium.
FAU - Van Eynde, A
AU  - Van Eynde A
FAU - Vulsteke, V
AU  - Vulsteke V
FAU - Connor, J
AU  - Connor J
FAU - Shenolikar, S
AU  - Shenolikar S
FAU - Stalmans, W
AU  - Stalmans W
FAU - Bollen, M
AU  - Bollen M
LA  - eng
GR  - DK52044/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Carrier Proteins)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (protein phosphatase inhibitor-1)
RN  - 452VLY9402 (Serine)
RN  - EC 3.1.- (Endoribonucleases)
RN  - EC 3.1.3.16 (Phosphoprotein Phosphatases)
RN  - EC 3.1.3.16 (Protein Phosphatase 1)
RN  - EC 3.1.4.- (PPP1R8 protein, human)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding, Competitive
MH  - *Carrier Proteins
MH  - Catalytic Domain
MH  - Cattle
MH  - Cell Nucleus/enzymology
MH  - *Endoribonucleases
MH  - Enzyme Inhibitors/*metabolism
MH  - Escherichia coli
MH  - Humans
MH  - *Intracellular Signaling Peptides and Proteins
MH  - Molecular Sequence Data
MH  - Muscle, Skeletal/enzymology
MH  - Peptide Mapping
MH  - Phosphoprotein Phosphatases/*metabolism
MH  - Phosphorylation
MH  - Protein Phosphatase 1
MH  - RNA-Binding Proteins/*metabolism
MH  - Rabbits
MH  - Recombinant Proteins/metabolism
MH  - Serine/metabolism
MH  - Spodoptera
MH  - Structure-Activity Relationship
MH  - Yeasts
EDAT- 1999/05/13 00:00
MHDA- 1999/05/13 00:01
CRDT- 1999/05/13 00:00
PHST- 1999/05/13 00:00 [pubmed]
PHST- 1999/05/13 00:01 [medline]
PHST- 1999/05/13 00:00 [entrez]
AID - 10.1074/jbc.274.20.14053 [doi]
AID - S0021-9258(19)73271-2 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 May 14;274(20):14053-61. doi: 10.1074/jbc.274.20.14053.