通过厌氧高效液相色谱法证明脱氧血红蛋白和碳氧血红蛋白之间的中间体

Demonstration of intermediates between deoxy and carbonmonoxy hemoglobins by anaerobic high-performance liquid chromatography.

作者信息

Anbari M, Asakura T

出版信息

FEBS Lett. 1986 Oct 6;206(2):253-6. doi: 10.1016/0014-5793(86)80991-7.

Abstract

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.

摘要

在室温下，使用配备阳离子交换柱的厌氧高效液相色谱系统分离与一氧化碳（CO）部分结合的中间亚铁血红蛋白（Hb）。通过将各种氧合血红蛋白和一氧化碳血红蛋白A的混合物应用于色谱柱来实现分离。流动相含有3 mM连二亚硫酸钠，它使氧合血红素脱氧，但使一氧化碳血红素保持完整，从而产生与不同数量CO分子结合的血红蛋白。对洗脱液的分光光度分析表明，血红蛋白按脱氧血红蛋白、Hb(CO)、Hb(CO)2、Hb(CO)3和Hb(CO)4的顺序洗脱。