Photochemical localization of the semotiadil binding region within the cardiac Ca2+ channel α1 subunit. Comparison with the skeletal muscle counterpart

FEBS Letters

Shibano, Toshiro¹  Ii, Nariaki²  Kawahara, Kohichi²  Kuniyasu, Akihiko²  Schwartz, Arnold³  Nakayama, Hitoshi² 

[1] New Product Research Laboratories II, Daiichi Pharmaceutical Co. Ltd., 1-16-13, Kita-Kasai, Edogawa-Ku, Tokyo 134-0081, Japan;Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Ohe Honmachi, Kumamoto 082-0973, Japan;Insitiute for Molecular Pharmacology and Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0828, USA

关键词: Semotiadil;    Binding site;    Calcium channel;    Cardiac muscle;    Photoaffinity labeling;   

DOI  :  10.1016/S0014-5793(98)01536-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have previously identified the binding region of a new Ca²⁺ antagonist semotiadil in the skeletal muscle Ca²⁺ channel. To the same semotiadil derivatives, the cardiac counterpart showed distinct and different binding characteristics: semotiadil and its photoaffinity analog D51-4700 inhibited [³H]PN200-110 binding to cardiac membrane preparations with IC₅₀ values of 13–20 μM, which are 10 times higher than those in skeletal muscle. Hill slopes of the binding inhibition were 0.94–1.0 for the cardiac channels compared to 0.63–0.67 for the skeletal muscle channels. A possible explanation for the difference is that the semotiadil binding site is differently conferred in cardiac and skeletal muscle Ca²⁺ channels. To reveal this within the primary structure, photoaffinity labeling of cardiac membranes was employed. [³H]D51-4700 was photoincorporated in several polypeptides but only the α1 subunit of the Ca²⁺ channel was photolabeled in a specific manner. Antibody mapping of the [³H]D51-4700-labeled α1 subunit with several anti-peptide antibodies revealed that the labeled site was located solely in a peptide fragment between Cys¹⁴⁶¹ and Lys¹⁵²⁹. This region encompasses the labeled site of skeletal muscle, but contains several non-identical amino acid residues, which may participate in expressing different binding characteristics between the two muscle type Ca²⁺ channels.

【 授权许可】

Unknown