A new synthetic substrate for protein methionine sulfoxide reductase is proposed. We show that extracts from human polymorphonuclear leukocytes can reduce 4-dimethylaminoazobenzene-4'-sulfonyl-L-methionine-dl-sulfoxide [DABS-L-Met-dl-(O)] to the corresponding methionine derivative, in the presence of dithiothreitol or dithioerythritol. The product of the reaction (DABS-Met) was separated by reversed-phase HPLC and detected by reading the absorbance at 436 nm. Due to the chirality of the sulfur atom in the sulfoxide, two diastereomers of Met(O) exist, namely Met-l-sulfoxide and Met-d-sulfoxide. After separation of the two forms and preparation of the DABS-derivatives, we observed a preferential reduction of the l-sulfoxide by polymorphonuclear leukocytes extracts. We discuss the possibility that the observed stereospecificity might have physiological relevance in the field of the oxidative modifications of proteins.
Reduction of DABS-L-methionine-dl-sulfoxide by protein methionine sulfoxide reductase from polymorphonuclear leukocytes: stereospecificity towards the l-sulfoxide
MINETTI, GIAMPAOLO;BALDUINI, CESARE;BROVELLI, AUGUSTA
1994-01-01
Abstract
A new synthetic substrate for protein methionine sulfoxide reductase is proposed. We show that extracts from human polymorphonuclear leukocytes can reduce 4-dimethylaminoazobenzene-4'-sulfonyl-L-methionine-dl-sulfoxide [DABS-L-Met-dl-(O)] to the corresponding methionine derivative, in the presence of dithiothreitol or dithioerythritol. The product of the reaction (DABS-Met) was separated by reversed-phase HPLC and detected by reading the absorbance at 436 nm. Due to the chirality of the sulfur atom in the sulfoxide, two diastereomers of Met(O) exist, namely Met-l-sulfoxide and Met-d-sulfoxide. After separation of the two forms and preparation of the DABS-derivatives, we observed a preferential reduction of the l-sulfoxide by polymorphonuclear leukocytes extracts. We discuss the possibility that the observed stereospecificity might have physiological relevance in the field of the oxidative modifications of proteins.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.