Atypical Reactive Center Kunitz-Type Inhibitor from the Sea Anemone Heteractis crispa

Marine Drugs

Irina Gladkikh³  Margarita Monastyrnaya³  Elena Leychenko³  Elena Zelepuga³  Victoria Chausova³  Marina Isaeva³  Stanislav Anastyuk³  Yaroslav Andreev¹  Steve Peigneur²  Jan Tytgat² 

[1] Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10, Miklukho-Maklaya Str., Moscow 117997, Russian Federation;Laboratory of Toxicology, University of Leuven (K.U. Leuven), Campus Gasthuisberg O&N2, Herestraat 49, P.O. Box 922, Leuven B-3000, Belgium;G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 159, Pr. 100 let Vladivostoku, Vladivostok 690022, Russian Federation;

关键词: sea anemone;    Kunitz-type protease inhibitor;    structure;    function;    channels;   

DOI  :  10.3390/md10071545

来源: mdpi

【 摘 要 】

The primary structure of a new Kunitz-type protease inhibitor InhVJ from the sea anemone Heteractis crispa (Radianthus macrodactylus) was determined by protein sequencing and cDNA cloning. InhVJ amino acid sequence was shown to share high sequence identity (up to 98%) with the other known Kunitz-type sea anemones sequences. It was determined that the P1 Thr at the reactive site resulted in a decrease of the K_i of InhVJ to trypsin and α-chymotrypsin (7.38 × 10⁻⁸ M and 9.93 × 10⁻⁷ M, respectively). By structure modeling the functional importance of amino acids at the reactive site as well as at the weak contact site were determined. The significant role of Glu45 for the orientation and stabilization of the InhVJ-trypsin complex was elucidated. We can suggest that there has been an adaptive evolution of the P1 residue at the inhibitor reactive site providing specialization or functional diversification of the paralogs. The appearance of a key so-called P1 Thr residue instead of Lys might lead to refinement of inhibitor specificity in the direction of subfamilies of serine proteases. The absence of Kv channel and TRPV1-receptor modulation activity was confirmed by electrophysiological screening tests.

【 授权许可】

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© 2012 by the authors; licensee MDPI, Basel, Switzerland.