Csk-homologous kinase interacts with SHPS-1 and enhances neurite outgrowth of PC12 cells

Hiroaki Mitsuhashi, Eugene Futai, Noboru Sasagawa, Yukiko Hayashi, Ichizo Nishino, Shoichi Ishiura

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)


SHPS-1 is an immunoglobulin superfamily protein with four immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in its cytoplasmic region. Various neurotrophic factors induce the tyrosine phosphorylation of SHPS-1 and the association of SHPS-1 with the protein tyrosine phosphatase SHP-2. Using a yeast two-hybrid screen, we identified a protein tyrosine kinase, Csk-homologous kinase (CHK), as an SHPS-1-interacting protein. Immunoprecipitation and pull-down assays using glutathione S-transferase (GST) fusion proteins containing the Src homology 2 (SH2) domain of CHK revealed that CHK associates with tyrosine-phosphorylated SHPS-1 via its SH2 domain. HIS3 assay in a yeast two-hybrid system using the tyrosine-to-phenylalanine mutants of SHPS-1 indicated that the first and second ITIMs of SHPS-1 are required to bind CHK. Over-expression of wild-type CHK, but not a kinase-inactive CHK mutant, enhanced the phosphorylation of SHPS-1 and its subsequent association with SHP-2. CHK phosphorylated each of four tyrosines in the cytoplasmic region of SHPS-1 in vitro. Co-expression of SHPS-1 and CHK enhanced neurite outgrowth in PC12 cells. Thus, CHK phosphorylates and associates with SHPS-1 and is involved in neural differentiation via SHP-2 activation.

Original languageEnglish
Pages (from-to)101-112
Number of pages12
JournalJournal of Neurochemistry
Issue number1
Publication statusPublished - 2008 Apr


  • CHK
  • Neurite
  • PC12
  • SHP-2
  • SHPS-1
  • Tyrosine phosphorylation


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