Dean J, Schechter A N
J Biol Chem. 1979 Sep 25;254(18):9185-93.
An anti-hemoglobin antiserum obtained from a sheep immunized with human carboxyhemoglobin A0 demonstrated little difference in its reactivity with deoxy- or carboxyhemoglobin A0. However, a subpopulation of this antiserum isolated by synthetic peptide affinity chromatography clearly distinguished between these two hemoglobin species. This subpopulation, designated alpha(129-141) anti-hemoglobin antibodies, represents less than 1% of the total anti-hemoglobin antibodies. They are nonprecipitating by Ouchterlony analysis, and fluorescence-quenching studies demonstrate the interaction of a single antibody binding site per hemoglobin dimer. These antibodies bind preferentially to carboxyhemoglobin with a median affinity constant of 5 X 10(8) M-1 compared to binding to deoxyhemoglobin with a binding affinity of less than 1 X 10(8) M-1. Furthermore, the presence of these antibodies in stoichiometric amounts increases the oxygen affinity of hemoglobin, and thus antibody and oxygen binding to hemoglobin can be considered as a linked function.
用人类羧基血红蛋白A0免疫绵羊获得的抗血红蛋白抗血清，在与脱氧血红蛋白A0或羧基血红蛋白A0的反应性上几乎没有差异。然而，通过合成肽亲和色谱法分离出的该抗血清亚群能清楚地区分这两种血红蛋白。这个亚群被命名为α(129 - 141)抗血红蛋白抗体，占总抗血红蛋白抗体的比例不到1%。通过双向免疫扩散分析它们不会沉淀，荧光猝灭研究表明每个血红蛋白二聚体有一个抗体结合位点相互作用。这些抗体优先结合羧基血红蛋白，中位亲和常数为5×10⁸ M⁻¹，而与脱氧血红蛋白结合的亲和常数小于1×10⁸ M⁻¹。此外，化学计量的这些抗体的存在会增加血红蛋白的氧亲和力，因此抗体与血红蛋白的结合以及氧与血红蛋白的结合可被视为一种关联功能。
