PMID- 10194355
OWN - NLM
STAT- MEDLINE
DCOM- 19990429
LR  - 20220309
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 14
DP  - 1999 Apr 6
TI  - Characterization of the neuronal targeting protein spinophilin and its
      interactions with protein phosphatase-1.
PG  - 4365-73
AB  - Protein phosphatase-1 (PP1) plays an important role in a variety of cellular
      processes, including muscle contraction, cell-cycle progression, and
      neurotransmission. The localization and substrate specificity of PP1 are
      determined by a class of proteins known as targeting subunits. In the present
      study, the interaction between PP1 and spinophilin, a neuronal protein that
      targets PP1 to dendritic spines, has been characterized. Deletion analysis
      revealed that a high-affinity binding domain is located within residues 417-494
      of spinophilin. This domain contains a pentapeptide motif (R/K-R/K-V/I-X-F)
      between amino acids 447 and 451 (R-K-I-H-F) that is conserved in other PP1
      regulatory subunits. Mutation of phenylalanine-451 (F451A) or deletion of the
      conserved motif abolished the ability of spinophilin to bind PP1, as observed by 
      coprecipitation, overlay, and competition binding assays. In addition, deletion
      of regions 417-442 or 474-494, either singly or in combination, impaired the
      ability of spinophilin to coprecipitate PP1. A comparison of the binding and
      inhibitory properties of spinophilin peptides suggested that distinct subdomains 
      of spinophilin are responsible for binding and modulating PP1 activity.
      Mutational analysis of the modulatory subdomain revealed that spinophilin
      interacts with PP1 via a mechanism unlike those used by the cytosolic inhibitors 
      DARPP-32 (dopamine- and cAMP-regulated phosphoprotein, Mr 32 000) and
      inhibitor-1. Finally, characterization of the interactions between spinophilin
      and PP1 has facilitated the design of peptide antagonists capable of disrupting
      spinophilin-PP1 interactions. These studies support the notion that spinophilin
      functions in vivo as a neuronal PP1 targeting subunit by directing the enzyme to 
      postsynaptic densities and regulating its activity toward physiological
      substrates.
FAU - Hsieh-Wilson, L C
AU  - Hsieh-Wilson LC
AD  - Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University,
      New York 10021, USA. hsiehl@rockvax.rockefeller.edu
FAU - Allen, P B
AU  - Allen PB
FAU - Watanabe, T
AU  - Watanabe T
FAU - Nairn, A C
AU  - Nairn AC
FAU - Greengard, P
AU  - Greengard P
LA  - eng
GR  - P01 DA010044/DA/NIDA NIH HHS/United States
GR  - DA10044/DA/NIDA NIH HHS/United States
GR  - MH40899/MH/NIMH NIH HHS/United States
PT  - Comparative Study
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  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Dopamine and cAMP-Regulated Phosphoprotein 32)
RN  - 0 (Microfilament Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Peptides)
RN  - 0 (Phosphoproteins)
RN  - 0 (Proteins)
RN  - 0 (neurabin)
RN  - 0 (phosphoprotein phosphatase inhibitor 1)
RN  - EC 3.1.3.16 (Phosphoprotein Phosphatases)
RN  - EC 3.1.3.16 (Protein Phosphatase 1)
SB  - IM
MH  - Animals
MH  - Binding Sites/drug effects
MH  - Cell Line
MH  - Dopamine and cAMP-Regulated Phosphoprotein 32
MH  - Humans
MH  - Microfilament Proteins/antagonists & inhibitors/*chemistry/metabolism
MH  - Nerve Tissue Proteins/chemistry
MH  - Neurons/enzymology/*metabolism
MH  - Peptide Fragments/metabolism
MH  - Peptides/chemical synthesis/pharmacology
MH  - Phosphoprotein Phosphatases/antagonists & inhibitors/*metabolism
MH  - *Phosphoproteins
MH  - Protein Phosphatase 1
MH  - Protein Structure, Tertiary
MH  - Proteins/chemistry
MH  - Rabbits
MH  - Sequence Homology, Amino Acid
EDAT- 1999/04/09 02:02
MHDA- 2000/03/11 09:00
CRDT- 1999/04/09 02:02
PHST- 1999/04/09 02:02 [pubmed]
PHST- 2000/03/11 09:00 [medline]
PHST- 1999/04/09 02:02 [entrez]
AID - 10.1021/bi982900m [doi]
AID - bi982900m [pii]
PST - ppublish
SO  - Biochemistry. 1999 Apr 6;38(14):4365-73. doi: 10.1021/bi982900m.