Purification and characterization of proteinase inhibitors from wild soja (Glycine soja) seeds.
Nine proteinase inhibitors, I-VIIa, VIIb, and VIII, were isolated from wild soja seeds by ammonium sulfate fractionation and successive chromatographies on SP-Toyopearl 650M, Sephacryl S-200SF, and DEAE-Toyopearl 650S columns. Reverse-phase HPLC finally gave pure inhibitors. All of the inhibitors inhibited trypsin with dissociation constants of 3.2-6.2 x 10(-9) M. Some of the inhibitors inhibited chymotrypsin and elastase as well. Two inhibitors (VIIb and VIII) with a molecular weight of 20,000 were classified as a soybean Kunitz inhibitor family. Others (I-VIla) had a molecular weight of about 8,000, and were stable to heat and extreme pH, suggesting that these belonged to the Bowman-Birk inhibitor family. Partial amino acid sequences of four inhibitors were also analyzed. The complete sequence of inhibitor IV was ascertained from the nucleotide sequences of cDNA clones encoding isoinhibitors homologous to soybean C-II.
Deshimaru
Masanobu
M
Department of Chemistry, Faculty of Science, Fukuoka University, Japan.
Hanamoto
Ryuji
R
Kusano
Chiho
C
Yoshimi
Shingo
S
Terada
Shigeyuki
S
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000378:metabolism; D007887:Fabaceae / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification* / Q000494:pharmacology*; D012639:Seeds / Q000737:chemistry*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000494:pharmacology
D011480:Protease Inhibitors; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D010196:Pancreatic Elastase; D014357:Trypsin