title: Glutathione mixed disulfides and heterogeneity of chicken hemoglobins.
abstract: 1. Adult chicken hemoglobins Hb A and Hb D interact with glutathione disulfide, GSSG. The major hemoglobin, Hb A, forms at least two new components, termed GHb AI and GHb AII, and Hb D forms at least one, GHb DI. 2. At pH 8.0 and 5 degrees C, glutathione disulfide (GSSG) in a molar excess of 50 x took 6 days to complete the reaction, although at pH 8.6 and 41 degrees C only 1 hr was needed, where the hemoglobins Hb A and Hb D were converted to their most mobile forms GHb AII and GHb DI. 3. Slight molar excess (2.7 GSSG/Hb, pH 7.4, 41 degrees C), reacting for 1 hr, showed extensive formation of GHb AI and some GHb AII. 4. Electrophoretic patterns, from the reaction products of 54 GSSG/Hb excess at different times, showed a marked pH dependence. 5. Titration with pCMB (p-chloromercuribezoic acid) of DTE (dithioerythrytol)-reduced samples showed 8.0 +/- 0.4 (N = 5) -SH (sulfhydryl) per tetramer. In hemolysates not reacted with DTE, 6.0 +/- 0.4 (N = 3) -SH were detected. 6. DTE-reduced and GSSG-reacted hemoglobins showed 4.6 +/- 0.5 (N = 7) -SH and 1.5 +/- 0.4 (N = 6) -SH, respectively, as titrated by DTNB, pH 8.0. DTE-reduced hemoglobins showed four fast-reacting -SH groups, no longer present in GSSG-reacted hemoglobins. 7. Our data indicate that chicken GHb AI and GHb DI probably have two glutathionyl residues per tetramer whereas GHb AII has four.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Departamento de Biofisica, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.
authors.forename: E
authors.initials: E
authors.lastname: Reischl
authors.forename: A L
authors.initials: AL
authors.lastname: Dafré
mesh_terms: D000818:Animals; D002645:Chickens / Q000097:blood*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000096:biosynthesis; D019803:Glutathione Disulfide; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism; D007526:Isoelectric Point
chemical_list: D006454:Hemoglobins; D005978:Glutathione; D019803:Glutathione Disulfide