The pro-oxidant role of protein SH groups of hemoglobin in rat erythrocytes exposed to menadione.
Menadione is selectively toxic to erythrocytes. Although GSH is considered a primary target of menadione, intraerythrocyte thiolic alterations consequent to menadione exposure are only partially known. In this study alterations of GSH and protein thiols (PSH) and their relationship with methemoglobin formation were investigated in human and rat red blood cells (RBC) exposed to menadione. In both erythrocyte types, menadione caused a marked increase in methemoglobin associated with GSH depletion and increased oxygen consumption. However, in human RBC, GSH formed a conjugate with menadione, whereas, in rat RBC it was converted to GSSG, concomitantly with a loss of protein thiols (corresponding to menadione arylation), and an increase in glutathione-protein mixed disulfides (GS-SP). Such differences were related to the presence of highly reactive cysteines, which characterize rat hemoglobin (cys beta125). In spite of the greater thiol oxidation in rat than in human RBC, methemoglobin formation and the rate of oxygen consumption elicited by menadione in both species were rather similar. Moreover, in repeated experiments under N2 or CO-blocked heme, it was found that menadione conjugation (arylation) in both species was not dependent on the presence of oxygen or the status of heme. Therefore, we assumed that GSH (human RBC) and protein (rat RBC) arylation was equally responsible for increased oxygen consumption and Hb oxidation. Moreover, thiol oxidation of rat RBC was strictly related to methemoglobin formation.
Lusini
Lorenzo
L
Department of Neuroscience, Pharmacology Unit, University of Siena, Via A. Moro 4, 53100 Siena, Italy.
Rossi
Ranieri
R
Giustarini
Daniela
D
Simplicio
Paolo Di
PD
D000818:Animals; D002248:Carbon Monoxide / Q000494:pharmacology; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008706:Methemoglobin / Q000096:biosynthesis; D010100:Oxygen / Q000378:metabolism; D010101:Oxygen Consumption; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000097:blood*; D013438:Sulfhydryl Compounds / Q000097:blood*; D024483:Vitamin K 3 / Q000494:pharmacology*
D011506:Proteins; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D024483:Vitamin K 3; D002248:Carbon Monoxide; D008706:Methemoglobin; D010100:Oxygen