A serine/threonine kinase which causes apoptosis-like cell death interacts with a calcineurin B-like protein capable of binding Na+/H+ exchanger1

Miho Matsumoto, Yoshihide Miyake, Mana Nagita, Hiroki Inoue, Daiya Shitakubo, Koji Takemoto, Chie Ohtsuka, Hiroshi Murakami, Norihiro Nakamura, Hiroshi Kanazawa

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

We surveyed proteins capable of binding to the cytoplasmic domain of Na+/H+ exchanger (NHE) 1 in a rat brain cDNA library with the yeast two-hybrid system. One clone obtained coded for a protein reported previously as a human calcineurin homologous protein (CHP). Since CHP is homologous to the regulatory subunit B of calcineurin, we expected a possible interacting partner of CHP like the catalytic subunit of calcineurin (calcineurin A), and surveyed this putative partner again with the yeast two-hybrid system. A clone thus obtained coded for a kinase, which is basically the same as that reported for human DRAK2. Overexpression of the rat homologue of DRAK2 caused apoptosis-like cell death of NIH3T3 cells, which was dependent on the kinase activity, confirming the previous result for DRAK2. The purified CHP and rat DRAK2 proteins synthesized in Escherichia coli could bind in vitro. CHP and rat DRAK2 expressed in COS-7 cells were found to be localized in the Golgi apparatus and nucleus, respectively. Some of them was also found in the membrane peripheral region. When they were coexpressed in the same cells, most of CHP moved to the nucleus where rat DRAK2 is located, suggesting in vivo interaction of these proteins. However, minor but significant fractions of both proteins were also found in the membrane peripheral region. Rat DRAK2 is expressed highly in thymus, spleen, and testis, where the apoptosis plays an important role in physiology.

Original languageEnglish
Pages (from-to)217-225
Number of pages9
JournalJournal of biochemistry
Volume130
Issue number2
DOIs
Publication statusPublished - Aug 2001

Keywords

  • Apoptosis
  • Calcineurin B-like protein
  • NHE
  • Serine/threonine kinase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'A serine/threonine kinase which causes apoptosis-like cell death interacts with a calcineurin B-like protein capable of binding Na<sup>+</sup>/H<sup>+</sup> exchanger<sup>1</sup>'. Together they form a unique fingerprint.

Cite this