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Demonstration of intermediates between deoxy and carbonmonoxy hemoglobins by anaerobic high‐performance liquid chromatography
Author(s) -
Anbari Mazen,
Asakura Toshio
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)80991-7
Subject(s) - sodium dithionite , chemistry , carbon monoxide , hemoglobin , elution , chromatography , dithionite , ferrous , column chromatography , high performance liquid chromatography , hemeprotein , heme , inorganic chemistry , organic chemistry , catalysis , enzyme
Intermediary ferrous hemoglobins (Hb) partially liganded with carbon monoxide (CO) were separated using an anaerobic high‐performance liquid chromatography system equipped with a cation‐exchange column at room temperature. Separations were achieved by applying various mixtures of oxy‐ and CO‐Hb A to the column. The mobile phase contained 3 mM sodium dithionite which deoxygenated the oxy hemes but left the CO hemes intact, thereby producing hemoglobins bound with various numbers of CO molecules. Spectrophotometric analyses of the eluent showed that hemoglobins were eluted in the order of deoxy hemoglobin, Hb(CO), Hb(CO) 2 , Hb(CO) 3 and Hb(CO) 4 .