Peptidase inhibitors from the salivary glands of the cockroach Nauphoeta cinerea |
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| Authors: | Yuliya Taranushenko Konstantin S Vinokurov Barbara Kludkiewicz Dalibor Kodrík Franti?ek Sehnal |
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| Institution: | 1. Biology Centre ASCR, Institute of Entomology, Brani?ovská 31, 370 05 ?eské Budějovice, Czech Republic;2. University of South Bohemia, Faculty of Science, Brani?ovská 31, 370 05 ?eské Budějovice, Czech Republic;1. Department of Molecular Medicine and Tissue Injury Defense Research Center, Ewha Womans University Medical School, Seoul, Republic of Korea;2. Department of Biochemistry, Ewha Womans University Medical School, Seoul, Republic of Korea;3. Department of Life and Nanopharmaceutical Sciences, College of Pharmacy, Kyung Hee University, Seoul, Republic of Korea;1. Tianjin Key Laboratory of Animal and Plant Resistance, School of Life Sciences, Tianjin Normal University, Tianjin, 300387, PR China;2. Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, 300071, PR China;3. State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, PR China;1. Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland;2. In Vitro and Cell Biotechnology Laboratory, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland;3. Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland;4. Department of Poultry Science, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland;1. Laboratory of Analysis, Treatment and Valorization of Pollutants of the Environment and Products (LATVPEP), Faculty of Pharmacy, University of Monastir, Monastir 5000, Tunisia;2. Laboratory of Transmissible Diseases and Biologically Active Substances, Tunisia;1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;3. Ningbo University, Ningbo 315211, China;4. SUN YAT-SEN University, Guangzhou 510275, China |
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| Abstract: | Inhibitory activity against subtilisin, proteinase K, chymotrypsin and trypsin was detected in the salivary glands and saliva of the cockroach Nauphoeta cinerea (Blattoptera: Blaberidae). Fractionation of the salivary glands extract by affinity chromatography followed by reverse-phase HPLC yielded five subtilisin-inhibiting peptides with molecular masses ranging from 5 to 14 kDa. N-terminal sequences and subsequently full-length cDNAs of inhibitors designated NcPIa and NcPIb were obtained. The NcPIa cDNA contains 216 nucleotides and encodes a pre-peptide of 72 amino-acid residues of which 19 make up the signal peptide. The cDNA of NcPIb consists of 240 nucleotides and yields a putative secretory peptide of 80 amino-acid residues. Mature NcPIa (5906.6 Da, 53 residues) and NcPIb (6713.3 Da, 60 residues) are structurally similar (65.4% amino acid overlap) single-domain Kazal-type peptidase inhibitors. NcPIa with Arg in P1 position and typical Kazal motif VCGSD interacted stoichiometrically (1:1) with subtilisin and was slightly less active against proteinase K. NcPIb with Leu in P1 and modified Kazal motif ICGSD had similar activity on subtilisin and no on proteinase K but was active on chymotrypsin. |
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