Selective oxidation of methionine residues in Kunitz-type protease inhibitors.
Bovine pancreatic trypsin inhibitor (BPTI, also known as aprotinin or Kunitz inhibitor, a mini-protein composed of 58 amino-acid residues, containing a single methionine residue at position 52) has been selectively oxidized by treatment with chloramine T, under mild conditions, to the methionyl sulfoxide derivative. Spleen inhibitor II (SI II, an isoform of BPTI containing two methionine residues at positions 18 and 52) has been oxidized under the same conditions. Oxidation affects the functional properties of the two inhibitors differently: the antiproteolytic activity of BPTI towards bovine trypsin and chymotrypsin, porcine kallikrein and human leukocyte elastase is not changed upon oxidation, while in the oxidized SI II, the affinity for both chymotrypsin and elastase decreases, with respect to the native protein. These results have been directly related to the oxidation of Met18 in SI II, located at the P'3 site in the contact area with the proteases.
Concetti
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Dipartimento di Biologia Cellulare, Università di Camerino.
Angeletti
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Fioretti
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Ascoli
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D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000032:analysis* / Q000302:isolation & purification; D002417:Cattle; D002700:Chloramines; D002850:Chromatography, Gel; D002853:Chromatography, Liquid; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D003488:Cyanogen Bromide; D006868:Hydrolysis; D008715:Methionine / Q000032:analysis*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D014105:Tosyl Compounds
D002700:Chloramines; D014105:Tosyl Compounds; C016300:chloramine-T; D007611:Aprotinin; D008715:Methionine; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D003488:Cyanogen Bromide