screening and cdna cloning of kv1 potassium channel toxins in sea anemones筛查和cdna克隆kv1钾通道在海葵毒素.pdfVIP

Mar. Drugs 2010, 8, 2893-2905; doi:10.3390/md8122893 OPEN ACCESS Marine Drugs ISSN 1660-3397 /journal/marinedrugs Article Screening and cDNA Cloning of Kv1 Potassium Channel Toxins in Sea Anemones Yoshikazu Yamaguchi 1, Yuichi Hasegawa 1, Tomohiro Honma 1,2, Yuji Nagashima 1 and Kazuo Shiomi 1,* 1 Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan-4, Minato-ku, Tokyo 108-8477, Japan 2 Department of Food Nutrition, Tokai University Junior College, Miyamae-cho, Aoi-ku, Shizuoka 420-8511, Japan * Author to whom correspondence should be addressed; E-Mail: shiomi@kaiyodai.ac.jp; Tel.: +81-3-5463-0601; Fax: +81-3-5463-0669. Received: 2 November 2010; in revised form: 19 November 2010 / Accepted: 1 December 2010 / Published: 2 December 2010 Abstract: When 21 species of sea anemones were screened for Kv1 potassium channel toxins by competitive inhibition of the binding of 125I-α-dendrotoxin to rat synaptosomal membranes, 11 species (two species of Actiniidae, one species of Hormathiidae, five species of Stichodactylidae and three species of Thalassianthidae) were found to be positive. Furthermore, full-length cDNAs encoding type 1 potassium channel toxins from three species of Stichodactylidae and three species of Thalassianthidae were cloned by a combination of RT-PCR, 3′RACE and 5′RACE. The precursors of these six toxins are commonly composed of signal peptide, propart and