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ACS Chem Neurosci. 2017 Jul 19;8(7):1554-1569. doi: 10.1021/acschemneuro.7b00076. Epub 2017 Apr 25.

Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity.

Author information

1
Department of Psychiatry, Icahn School of Medicine at Mount Sinai , New York, New York 10029, United States.

Abstract

Kalirin7 (Kal7), a postsynaptic Rho GDP/GTP exchange factor (RhoGEF), plays a crucial role in long-term potentiation and in the effects of cocaine on behavior and spine morphology. The KALRN gene has been linked to schizophrenia and other disorders of synaptic function. Mass spectrometry was used to quantify phosphorylation at 26 sites in Kal7 from individual adult rat nucleus accumbens and prefrontal cortex before and after exposure to acute or chronic cocaine. Region- and isoform-specific phosphorylation was observed along with region-specific effects of cocaine on Kal7 phosphorylation. Evaluation of the functional significance of multisite phosphorylation in a complex protein like Kalirin is difficult. With the identification of five tyrosine phosphorylation (pY) sites, a panel of 71 SH2 domains was screened, identifying subsets that interacted with multiple pY sites in Kal7. In addition to this type of reversible interaction, endoproteolytic cleavage by calpain plays an essential role in long-term potentiation. Calpain cleaved Kal7 at two sites, separating the N-terminal domain, which affects spine length, and the PDZ binding motif from the GEF domain. Mutations preventing phosphorylation did not affect calpain sensitivity or GEF activity; phosphomimetic mutations at specific sites altered protein stability, increased calpain sensitivity, and reduced GEF activity.

KEYWORDS:

RhoGEF; cocaine; mass spectrometry; nucleus accumbens; prefrontal cortex; tyrosine phosphorylation

PMID:
28418645
PMCID:
PMC5517348
DOI:
10.1021/acschemneuro.7b00076
[Indexed for MEDLINE]
Free PMC Article

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