Studies on soybean trypsin inhibitors, 2, amino-acid sequence around the ractive site of soybean trypsin inhibitor (Kunitz)

Studies on soybean trypsin inhibitors, 2, amino-acid sequence around the ractive site of soybean trypsin inhibitor (Kunitz)

1973

REP.SB-0652

For the elucidation of amino acid sequence around the reactive site of soybean trypsin inhibitor (Kunitz), fragments A and B were digested with trypsin, and the resulting peptides were separated by ion-exchange chromatography on Dowex 50X2 or by gel filtration on Bio-Gel P-4. Further purification of the peptides was carried out by gel filtration on Sephadex G-25 or by high-voltage paper electrophoresis at pH 3.6 and 6.5. Nine peptides were obtained in pure form from fragment A and three peptides from fragment B, and their amino acid sequences were determined by the direct Edman method and by carboxy-peptidase digestion technique. Overlapping peptides necessary for the alignment of the tryptic peptides from fragments A and B were obtained from a chymotryptic hydrolysate of fragment AB by gel filtration on Bio-Gel P-4, and their amino acid compositions and sequence analyses of some peptides made it possible to establish the amino acid sequence of an amino-terminal region of the inhibitor consisting of 84 amino acid residues. The reactive site (Arg63-Ile64) of the inhibitor to trypsin was involved in this region. [AS]

v.32:408-416, EUROPEAN JOURNAL OF BIOCHEMISTRY

Research > Published Articles