Oxidative modification of aldose reductase induced by copper ion. Factors and conditions affecting the process.
Bovine lens aldose reductase (ALR2) is inactivated by copper ion [Cu(II)] through an oxygen-independent oxidative modification process. A stoichiometry of 2 equiv of Cu(II)/enzyme mol is required to induce inactivation. While metal chelators such as EDTA or o-phenantroline prevent but do not reverse the ALR2 inactivation, DTT allows the enzyme activity to be rescued by inducing the recovery of the native enzyme form. The inactive enzyme form is characterized by the presence of 2 equiv of bound copper, at least one of which present as Cu(I), and by the presence of two lesser equivalents, with respect to the native enzyme, of reduced thiol residues. Data are presented which indicate that the Cu-induced protein modification responsible for the inactivation of ALR2 is the generation on the enzyme of an intramolecular disulfide bond. GSH significantly interferes with the Cu-dependent inactivation of ALR2 and induces, through its oxidation to GSSG, the generation of an enzyme form linked to a glutathionyl residue by a disulfide bond.
Dipartimento di Fisiologia e Biochimica, Università di Pisa, Italy.
I
I
Cecconi
M
M
Moroni
P G
PG
Vilardo
M
M
Dal Monte
P
P
Borella
G
G
Rastelli
L
L
Costantino
D
D
Garland
D
D
Carper
J M
JM
Petrash
A
A
Del Corso
U
U
Mura
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D000693:Anaerobiosis; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D003300:Copper / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D007908:Lens, Crystalline / Q000201:enzymology; D010100:Oxygen / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism / Q000494:pharmacology
D002413:Cations, Divalent; D013438:Sulfhydryl Compounds; D003300:Copper; D000449:Aldehyde Reductase; D005978:Glutathione; D010100:Oxygen