A calcium-binding protein from rabbit lung cytosol identified as the product of growth-regulated gene (2A9) and its binding proteins

Hiroshi Tokumitsu, Ryoji Kobayashi, Hiroyoshi Hidaka

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Using Ca2+-dependent affinity chromatography on a synthetic compound (W-77)-coupled Sepharose 4B column, we purified two different Ca2+-binding proteins from rabbit lung extracts. The molecular weights of these proteins were estimated to be 17 kDa (calmodulin) and 10 kDa, respectively. The partial amino acid sequence of the 10-kDa protein revealed that it has two EF-hand structures. In addition, the 10-kDa protein was highly homologous (91%) to the product of growth-regulated gene, 2A9 (calcyclin). The Ca2+-binding property of the 10-kDa protein was observed by a change in the uv difference spectrum. Equilibrium dialysis showed that 1 mol of the 10-kDa protein bound to 2.04 ± 0.05 mol of Ca2+ in the presence of 10-4 m Ca2+. However, the protein failed to activate calmodulin-dependent enzymes such as Ca2+ CaM kinase II, myosin light chain kinase, and phosphodiesterase. We found that a 50-kDa cytosolic protein of the rabbit lung, intestine, and spleen bound to the 10-kDa protein, in a Ca2+-dependent manner. The distribution of calcyclin and calcyclin binding proteins was unique and seems to differ from that of calmodulin and calmodulin-binding proteins. Thus, calcyclin probably plays a physiological role through its binding proteins for the Ca2+-dependent cellular response.

Original languageEnglish
Pages (from-to)202-207
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume288
Issue number1
DOIs
Publication statusPublished - 1991
Externally publishedYes

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Calcium-Binding Proteins
Cytosol
Carrier Proteins
Genes
Rabbits
Lung
Growth
Proteins
Calmodulin
Calmodulin-Binding Proteins
EF Hand Motifs
Myosin-Light-Chain Kinase
Affinity chromatography
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Dialysis
Phosphoric Diester Hydrolases
Affinity Chromatography
Sepharose
Intestines
Amino Acid Sequence

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A calcium-binding protein from rabbit lung cytosol identified as the product of growth-regulated gene (2A9) and its binding proteins. / Tokumitsu, Hiroshi; Kobayashi, Ryoji; Hidaka, Hiroyoshi.

In: Archives of Biochemistry and Biophysics, Vol. 288, No. 1, 1991, p. 202-207.

Research output: Contribution to journalArticle

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