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J. Biochem, 1990, Vol. 108, No. 2 261-266
© 1990 Japanese Biochemical Society

research-article

Tachyplesins Isolated from Hemocytes of Southeast Asian Horseshoe Crabs (Carcinoscorpius rotundicauda and Tachypleus gigas)

Identification of a New Tachyplesin, Tachyplesin III, and a Processing Intermediate of Its Precursor1

Tatsushi Muta*, Tamiji Fujimoto**, Hiroshi Nakajima*** and Sadaaki Iwanaga*,**,2

*Department of Molecular Biology, Graduate School of Medical Science, Kyushu Univeredy 33 Hzgashi-kn Fuhuoka, Fukuoka 812
**Department of Biology, Faculty of Science, Kyushu Univeredy 33 Hzgashi-kn Fuhuoka, Fukuoka 812
***Taiyo Central R & D Institude, Taiyo Fishery Co., Ltd. Chuo-ku, Tokyo 104

2To whom correspondence should be addressed

Tachyplesins and their analogs are antimicrobial peptides composed of 17 or 18 amino acid residues present abundantly in acid extracts of hemocyte debris of horseshoe crabs. We purified here tachyplesln isopeptides from hemocytes of two species of Southeast Asian horseshoe crabs, Carcinoscorpius rotundicauda and Tachypleus gigas, and determined t)leir amino acid sequences. The major tachyplesin Isolated from both species was identified, respectively, as tachyplesin I, which had previously been found in hemocytes of the Japanese horseshoe crab (Tachypleus tridentatus). The yield from both species was very high (more than 70 mg per 100 g wet weight of hemocytes), i.e., comparable with that from T. tridentatu.s. In addition to tachyplesin I, a new tachyplesin isopeptide, named tachyplesin III, was also isolated from T. gigas hemocytes, in which an arginine replaced the 15th lysine of tachyplesin I. The carboxyl-terminal residue of the isolated tachyplesins I and III was confirmed, respectively, to be an arginine {alpha}-amide by chemical analysis. Furthermore, a tachyplesin peptide derivative with a carboxyl-terminal extension of glycine-lysine was newly found in the hemocytes of C. rotundicauda. It appeared to be an intermediate derived from a tachyplesin precursor during processing to the mature form.

1Thi study was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and CuIttu of Japan.


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