Different types of glutathionylation of hemoglobin can exist in intact erythrocytes.
Glutathionylation of hemoglobin (Hb) was studied by incubation of intact human erythrocytes with 1 mM tert-butylhydroperoxide (tBHP). Electrophoresis of the membranes showed a time dependent increase of membrane-bound Hb alpha chain until 10 min, and immunoblotting study showed that membrane-bound Hb alpha chain reacted with anti-glutathione antibody only after 10 min. Concomitant with the Hb alpha chain, membrane associated actin, spectrin, and glyceraldehyde 3-phosphate dehydrogenase reacted with the antibody. Cytosolic Hb of the control erythrocytes reacted with anti-glutathione antibody. Together with our previous paper, the present study indicates that at least three different types of glutathionylation of Hb can exist in erythrocytes. The first type is a mixed disulfide bond between reduced glutathione (GSH) and normal Hb. The second type is a disulfide bond between the cysteine 93 of metHb beta chain and oxidized glutathione (GSSG), and the third type is a disulfide bond between the other cysteine residues of metHb alpha chain and/or metHb beta chain and GSSG.
Mawatari
Shiro
S
Department of Nutrition and Health Science, Faculty of Human Environmental Science, Fukouka Women's University, 1-1-1 Kasumigaoka, Higashi-ku, Fukuoka 813-8529, Japan. s-mawatari@fwu.ac.jp
Murakami
Kaori
K
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism; D019803:Glutathione Disulfide / Q000737:chemistry / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008623:Mercaptoethanol / Q000737:chemistry; D008706:Methemoglobin / Q000737:chemistry / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D013049:Spectrin / Q000737:chemistry / Q000378:metabolism; D020122:tert-Butylhydroperoxide / Q000737:chemistry / Q000494:pharmacology
D000199:Actins; D004220:Disulfides; D006454:Hemoglobins; D008565:Membrane Proteins; D013049:Spectrin; D008623:Mercaptoethanol; D008706:Methemoglobin; D020122:tert-Butylhydroperoxide; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005978:Glutathione; D003545:Cysteine; D019803:Glutathione Disulfide