caa a22 4500 397555725 CHVBK 20180308164801.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021549 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021549 Tachycitin, a Small Granular Component in Horseshoe Crab Hemocytes, Is an Antimicrobial Protein with Chitin-Binding Activity [Elektronische Daten] [Shun-ichiro Kawabata, Ranko Nagayama, Michimasa Hirata, Takeshi Shigenaga, Kishan Lal Agarwala, Tetsu Saito, Junko Cho, Hiroshi Nakajima, Toshio Takagi, Sadaaki Iwanaga] Small granules of horseshoe crab hemocytes contain two known major antimicrobial substances, tachyplesin and big defensin (S5), and at least five protein components (S1 58), with unknown functions. In the present study, we examined the biological properties and primary structure of a small granular component S2, named tachycitin. This component was purified from the acid extract of hemocyte debris by two steps of chromatography. The purified tachycitin was a single chain protein with an apparent Mr =8,500 on Tricine-SDS-polyacrylamide gel electrophoresis. Ultracentrifugation analysis revealed tachycitin to be present in monomer form in solution. Tachycitin inhibited the growth of both Gram-negative and -positive bacteria, and fungi, with a bacterial agglutinating property. Moreover, tachycitin and big defensin acted synergistically in antimicrobial activities. The amino acid sequence and intrachain disulfide bonds of tachycitin were determined by amino acid and sequence analyses of peptides produced by enzymatic cleavages. The mature tachycitin.achycitin consisted of 73 amino acid residues containing five disulfide bonds with no N-linked sugar. A cDNA coding for tachycitin was isolated from a hemocyte cDNA library. The open reading frame coded for an NH2-terminal signal sequence followed by the mature peptide and an extension sequence of -Gly-Arg-Lys at the COOH-terminus, which is a putative amidating signal. The COOH-terminal threonine amide released after digestion of tachycitin with lysylendopeptidase was identified. The NH2-terminal 28 residues tachycitin shows sequence homology to a part of chitin-binding regions found in antifungal chitin-binding peptides, chitin-binding lectins, and chitinases, all of which have been isolated from plants. Tachycitin showed a specific binding to chitin but did not bind with the polysaccharides cellulose, mannan, xylan, and laminarin. Tachycitin may represent new class of chitin-binding protein family in animals. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence agglutinin nationallicence antimirobial activity nationallicence chitin bindings nationallicence horseshoe crab nationallicence host defense nationallicence Kawabata Shun-ichiro Department of Biology, Faculty of Science Fukuoka 812-81 aut Nagayama Ranko Department of Biology, Faculty of Science Fukuoka 812-81 aut Hirata Michimasa Department of Bacteriology, School of Medicine, Iwate Medical University Morioka 020 aut Shigenaga Takeshi Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut Agarwala Kishan Lal Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut Saito Tetsu Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut Cho Junko Central Research Institute, Maruha Co., Ltd. Tsukuba 300-42 aut Nakajima Hiroshi Central Research Institute, Maruha Co., Ltd. Tsukuba 300-42 aut Takagi Toshio the Institute for Protein Research, Osaka University Suita, Osaka 565 aut Iwanaga Sadaaki Department of Biology, Faculty of Science Fukuoka 812-81 aut The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1253-1260 0021-924X 120:6<1253 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021549 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021549 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kawabata Shun-ichiro Department of Biology, Faculty of Science Fukuoka 812-81 aut NATIONALLICENCE 700 1- Nagayama Ranko Department of Biology, Faculty of Science Fukuoka 812-81 aut NATIONALLICENCE 700 1- Hirata Michimasa Department of Bacteriology, School of Medicine, Iwate Medical University Morioka 020 aut NATIONALLICENCE 700 1- Shigenaga Takeshi Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Agarwala Kishan Lal Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Saito Tetsu Department of Molecular Biology, Graduate School of Medical Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Cho Junko Central Research Institute, Maruha Co., Ltd. Tsukuba 300-42 aut NATIONALLICENCE 700 1- Nakajima Hiroshi Central Research Institute, Maruha Co., Ltd. Tsukuba 300-42 aut NATIONALLICENCE 700 1- Takagi Toshio the Institute for Protein Research, Osaka University Suita, Osaka 565 aut NATIONALLICENCE 700 1- Iwanaga Sadaaki Department of Biology, Faculty of Science Fukuoka 812-81 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1253-1260 0021-924X 120:6<1253 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford