Neurocalcin (NC), a neuron-specific EF-hand Ca2+-binding protein, purified from bovine brain [Terasawa, Nakano, Kobayashi and Hidaka (1992) J. Biol. Chem. 267, 19596–19599] contains multiple isoforms. We previously cloned NCδ from bovine brain and showed high expression in neuronal tissues [Okazaki, Watanabe, Ando, Hagiwara, Terasawa and Hidaka (1992) Biochem. Biophys. Res. Commun. 185, 147–153]. We report here the molecular cloning and expression of a cDNA encoding bovine brain NCα. The translated bovine protein is 191 amino acids long and shares 69.1% of its amino acid sequence with NCδ. Recombinant NCα migrates as a single 23 kDa band and exhibits a Ca2+-dependent mobility shift on SDS/PAGE. Analysis of fluorescence emission spectra showed the Ca2+-induced peak at 337 nm. Interestingly, the mobility shift and the fluorescence intensity at 337 nm were larger for NCα than for NCδ. In Ca2+-overlay experiments, however, the apparent affinity of NCα for 45Ca2+ was similar to that of NCδ. Immunohistochemical analysis revealed NCα expression in the granular layer of the rat cerebellar cortex whereas NCδ was found in the Purkinje cell layer. In the rat olfactory bulb, NCα was located in external tufted cells, and NCδ was found in the periglomerular cells. These data demonstrate that NC isoforms differ in their tissue distribution and conformational changes induced by Ca2+ binding. Thus differential regulation of the two NC isoforms may be involved in control of neuron function.
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Research Article|
May 01 1998
Cloning and expression of a cDNA encoding a new neurocalcin isoform (neurocalcin α) from bovine brain
Masumi KATO;
Masumi KATO
*Department of Pharmacology, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
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Yasuo WATANABE;
Yasuo WATANABE
*Department of Pharmacology, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
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Satoshi IINO;
Satoshi IINO
†Department of Anatomy, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
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Yuri TAKAOKA;
Yuri TAKAOKA
*Department of Pharmacology, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
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Shigeru KOBAYASHI;
Shigeru KOBAYASHI
†Department of Anatomy, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
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Tatsuya HAGA;
Tatsuya HAGA
‡Department of Biochemistry, Institute for Brain Research, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan
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Hiroyoshi HIDAKA
Hiroyoshi HIDAKA
1
*Department of Pharmacology, Nagoya University School of Medicine, Tsurumai 65, Showa-ku, Nagoya 466, Japan
1To whom correspondence should be addressed (e-mail hhidaka@tsuru.med.nagoya-u.ac.jp).
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Publisher: Portland Press Ltd
Received:
October 06 1997
Revision Received:
January 26 1998
Accepted:
February 10 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 331 (3): 871–876.
Article history
Received:
October 06 1997
Revision Received:
January 26 1998
Accepted:
February 10 1998
Citation
Masumi KATO, Yasuo WATANABE, Satoshi IINO, Yuri TAKAOKA, Shigeru KOBAYASHI, Tatsuya HAGA, Hiroyoshi HIDAKA; Cloning and expression of a cDNA encoding a new neurocalcin isoform (neurocalcin α) from bovine brain. Biochem J 1 May 1998; 331 (3): 871–876. doi: https://doi.org/10.1042/bj3310871
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