caa a22 4500 397556241 CHVBK 20180308164803.0 cr unu---uuuuu 161202e199606 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021355 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021355 Purification, Characterization, and Sequencing of Two Cysteine Proteinase Inhibitors, Sea and Scb, from Sunflower (Helianthus annuus) Seeds [Elektronische Daten] [Yoshiaki Kouzuma, Keiko Kawano, Makoto Kimura, Nobuyuki Yamasaki, Tomoko Kadowaki, Kenji Yamamoto] Two proteinaceous cysteine proteinase inhibitors (cystatins) referred to as Sea and Scb were purified to homogeneity from the seeds of sunflower (Heliantas annuus) by gel filtration on Sephadex G-75 followed by a series of ion-exchange column chromatographies and reverse-phase HPLC (RP-HPLC). The isoelectric points (pi) of Sea and Scb were estimated to be 5.6 and 9.6, respectively. The inhibitory potencies of these two cystatins were examined with cysteine proteinases from various sources, such as plants, mammals, and bacteria. Papain was strongly inhibited by both Sea and Scb with K1 values of 5.6 X10−9 and 1.7 × 10−10 M, respectively. Sea and Scb were also found to be potent inhibitors of ficin (K1 values of 1.9×10−6 and 2.8×10−9M, respectively). Rat cathepsin H was inhibited strongly by Scb and slightly by Sea. Although rat cathepsins B and L were significantly inhibited by Scb, they were scarcely affected by Sea. Neither Sea nor Scb inhibited Arg-gingipain, an arginine-specific cysteine proteinase of Porphyromonas gingivalis. The complete amino acid sequences of the two inhibitors were determined by protein chemical methods. The proteins Sea and Scb consist of 83 and 101 amino acid residues with Mr of 9,330 and 11,187, respectively, and there are identical residues at 34 positions in the two sequences, that is at 42% of the residues compared. Comparison of their sequences with those of other cystatins revealed that Sea shares 59-73% identical residues with other phytocystatins, while Scb shows less identity to other phytocystatins, sharing only 28-38% identical residues. Furthermore, only 20-27% of the residues of both cystatins, Sea and Scb, are identical to those of the animal cystatins. © by the Journal of Biochemistry Regular Papers nationallicence amino acid sequence nationallicence cysteine proteinase inhibitor nationallicence Helianthus annuus nationallicence phytocystatin nationallicence sunflower nationallicence Kouzuma Yoshiaki Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut Kawano Keiko Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut Kimura Makoto Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut Yamasaki Nobuyuki Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut Kadowaki Tomoko Department of Pharmacology, Faculty of Dentistry, Kyushu University Higashi-ku, Fukuoka 812 aut Yamamoto Kenji Department of Pharmacology, Faculty of Dentistry, Kyushu University Higashi-ku, Fukuoka 812 aut The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1106-1113 0021-924X 119:6<1106 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021355 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021355 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kouzuma Yoshiaki Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 700 1- Kawano Keiko Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 700 1- Kimura Makoto Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 700 1- Yamasaki Nobuyuki Laboratory of Biochemistry, Faculty of Agriculture Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 700 1- Kadowaki Tomoko Department of Pharmacology, Faculty of Dentistry, Kyushu University Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 700 1- Yamamoto Kenji Department of Pharmacology, Faculty of Dentistry, Kyushu University Higashi-ku, Fukuoka 812 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1106-1113 0021-924X 119:6<1106 1996 119 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