title: Thiol-dependent metal-catalyzed oxidation of bovine lens aldose reductase. II. Proteolytic susceptibility of the modified enzyme form.
abstract: Bovine lens aldose reductase (alditol: NADP+ oxidoreductase, EC 1.1.1.21) undergoes a modification induced by 2-mercaptoethanol in the presence of the redox system Fe(II)/Fe(III). The modified form (ARa) exhibits an increased hydrophobicity and tendency to aggregate. Moreover, while the native enzyme form is rather insensitive to proteolytic breakdown, the modified form is susceptible to limited proteolysis by trypsin and chymotrypsin. With both proteases, the degradation correlated with a loss of enzyme activity and results in the appearance of one molecular species of 26 KDa (for chymotrypsin) and two molecular species of 24 and 17 KDa (for trypsin). The decline in solubility and the increase in susceptibility to proteolysis of ARa suggests that the thiol-dependent metal-catalyzed modification is comparable to other oxidative systems that mark proteins for degradation.
authors.affiliation: Dipartimento di Fisiologia e Biochimica, Universitá di Pisa, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Del Corso
authors.forename: M
authors.initials: M
authors.lastname: Voltarelli
authors.forename: M
authors.initials: M
authors.lastname: Giannessi
authors.forename: M
authors.initials: M
authors.lastname: Cappiello
authors.forename: D
authors.initials: D
authors.lastname: Barsacchi
authors.forename: M
authors.initials: M
authors.lastname: Zandomeneghi
authors.forename: M
authors.initials: M
authors.lastname: Camici
authors.forename: U
authors.initials: U
authors.lastname: Mura
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D007501:Iron / Q000494:pharmacology*; D007700:Kinetics; D007908:Lens, Crystalline / Q000201:enzymology*; D008623:Mercaptoethanol / Q000494:pharmacology*; D008970:Molecular Weight; D009249:NADP / Q000494:pharmacology; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D014357:Trypsin / Q000494:pharmacology*
chemical_list: D010446:Peptide Fragments; D009249:NADP; D008623:Mercaptoethanol; D007501:Iron; D000449:Aldehyde Reductase; D002918:Chymotrypsin; D014357:Trypsin