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Toxins 2016, 8(7), 205; doi:10.3390/toxins8070205

Cloning a Chymotrypsin-Like 1 (CTRL-1) Protease cDNA from the Jellyfish Nemopilema nomurai

1
Department of Pharmacology and Toxicology, College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Korea
2
Marine Environment Research Center, Korea Institute of Ocean Science and Technology (KIOST), Geoje 656-834, Korea
3
Faculty of Marine Environmental Science, University of Science and Technology (UST), Geoje 656-834, Korea
4
Ballast Water Research Center, Korea Institute of Ocean Science and Technology (KIOST), Geoje 656-834, Korea
5
Headquarters for Marine Environment, National Fisheries Research & Development Institute, Shiran-ri, Gijang-eup, Gijang-gun, Busan 619-705, Korea
6
Institutes of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Korea
7
Engineering Research Institute, Gyeongsang National University, Jinju-si 660-701, Korea
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Meg Daly
Received: 16 March 2016 / Revised: 30 May 2016 / Accepted: 28 June 2016 / Published: 5 July 2016
(This article belongs to the Section Animal Venoms)
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Abstract

An enzyme in a nematocyst extract of the Nemopilema nomurai jellyfish, caught off the coast of the Republic of Korea, catalyzed the cleavage of chymotrypsin substrate in an amidolytic kinetic assay, and this activity was inhibited by the serine protease inhibitor, phenylmethanesulfonyl fluoride. We isolated the full-length cDNA sequence of this enzyme, which contains 850 nucleotides, with an open reading frame of 801 encoding 266 amino acids. A blast analysis of the deduced amino acid sequence showed 41% identity with human chymotrypsin-like (CTRL) and the CTRL-1 precursor. Therefore, we designated this enzyme N. nomurai CTRL-1. The primary structure of N. nomurai CTRL-1 includes a leader peptide and a highly conserved catalytic triad of His69, Asp117, and Ser216. The disulfide bonds of chymotrypsin and the substrate-binding sites are highly conserved compared with the CTRLs of other species, including mammalian species. Nemopilema nomurai CTRL-1 is evolutionarily more closely related to Actinopterygii than to Scyphozoan (Aurelia aurita) or Hydrozoan (Hydra vulgaris). The N. nomurai CTRL1 was amplified from the genomic DNA with PCR using specific primers designed based on the full-length cDNA, and then sequenced. The N. nomurai CTRL1 gene contains 2434 nucleotides and four distinct exons. The 5′ donor splice (GT) and 3′ acceptor splice sequences (AG) are wholly conserved. This is the first report of the CTRL1 gene and cDNA structures in the jellyfish N. nomurai. View Full-Text
Keywords: Nemopilema nomurai; amidolytic kinetic assay; cloning a chymotrypsin-like 1 (CTRL-1) protease; full-length cDNA sequence; genomic DNA sequence Nemopilema nomurai; amidolytic kinetic assay; cloning a chymotrypsin-like 1 (CTRL-1) protease; full-length cDNA sequence; genomic DNA sequence
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Heo, Y.; Kwon, Y.C.; Bae, S.K.; Hwang, D.; Yang, H.R.; Choudhary, I.; Lee, H.; Yum, S.; Shin, K.; Yoon, W.D.; Kang, C.; Kim, E. Cloning a Chymotrypsin-Like 1 (CTRL-1) Protease cDNA from the Jellyfish Nemopilema nomurai. Toxins 2016, 8, 205.

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