Specific modification of the functional arginine residue in soybean trypsin inhibitor (Kunitz) by peptidylarginine deiminase.
In order to elucidate the specificity of rabbit muscle peptidylarginine deiminase, which catalyzes the conversion of arginyl to citrullyl residues (Takahara, H., Oikawa, Y., and Sugawara, K. (1983) J. Biochem. (Tokyo) 94, 1945-1953), we examined the action of this enzyme on a variety of trypsin inhibitors by assay of residual trypsin-inhibiting activity. The enzyme rapidly abolished the activity of soybean trypsin inhibitor (Kunitz) (STI) in a process that was pseudo-first order with the rate dependent on enzyme concentration (second order rate constant = 5.0 X 10(4) M-1 S-1), whereas no detectable changes in activity were noted for other inhibitors tested. Inactivation of STI was due to the conversion of 1 arginine to a citrulline residue and was accompanied with a 0.2 unit decrease of the isoelectric point. There was no alteration of the molecular size and overall conformation of STI. Furthermore, analysis of modified STI indicated that arginine 63, known as the reactive site of STI, is the residue modified by peptidylarginine deiminase. Thus, peptidylarginine deiminase selectively catalyzes the deimination of the functional arginine residue of STI.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001120:Arginine / Q000378:metabolism*; D001665:Binding Sites; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000378:metabolism*; D007525:Isoelectric Focusing; D007700:Kinetics; D008433:Mathematics; D010059:Ovomucin / Q000378:metabolism; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000378:metabolism
D000596:Amino Acids; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; D010059:Ovomucin; D014360:Trypsin Inhibitor, Kunitz Soybean; D001120:Arginine; D006867:Hydrolases; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases