McGarrity M J, Er S S, Hsia J C
Biosciences Division, Defence and Civil Institute of Environmental Medicine, Downsview, Ontario, Canada.
J Chromatogr. 1987 Aug 7;419:37-50. doi: 10.1016/0378-4347(87)80264-5.
Six fractions (I-VI) of pyridoxal 5'-phosphate (PLP) hemoglobin (Hb), prepared by the method of De Venuto and Zegna [J. Surg. Res., 34 (1983) 205] have been isolated and purified by anion-exchange high-performance liquid chromatography (HPLC). Total phosphate analyses indicate that I is HbA, II and III are double-labelled, IV and V are tetra-labelled and VI contains 6 mol of phosphorus per mol of hemoglobin. The purified components have been resolved into their alpha and beta chains by preparative reversed-phase HPLC using a macroporous C4 support. Phosphate analyses indicate that the beta chains of II, III and IV each contain one phosphate per chain while the beta chains of V and VI each contain two phosphates. The alpha chains of IV and VI were found to be monophosphate-labelled. Reversed phase HPLC analysis of the tryptic peptides of the beta chains indicates that the label is bound exclusively to the 1-valine residue in II, III and IV while both the 1-valine and the 82-lysine are labelled in V and VI. Similarly, modification of the 1-valine residue of the alpha chains of IV and VI was detected. Components II and III have the same molecular formula. Evidence is presented which shows that they are interconvertible and that they correspond to the PLP2-Hb species and component V is Benesch's PLP4-Hb [J. Biol. Chem., 257 (1983) 1320 and references cited therein]. Component IV is III with one additional PLP per alpha chain and similarly VI is V with monolabelled alpha chains.(ABSTRACT TRUNCATED AT 250 WORDS)
采用德韦努托和泽尼亚的方法[《外科研究杂志》，34(1983)205]制备的6个磷酸吡哆醛(PLP)血红蛋白(Hb)级分(I - VI)，已通过阴离子交换高效液相色谱(HPLC)进行了分离和纯化。总磷酸盐分析表明，I是HbA，II和III是双标记的，IV和V是四标记的，VI每摩尔血红蛋白含有6摩尔磷。使用大孔C4载体，通过制备型反相HPLC将纯化后的组分分离为其α链和β链。磷酸盐分析表明，II、III和IV的β链每条含有一个磷酸盐，而V和VI的β链每条含有两个磷酸盐。发现IV和VI的α链是单磷酸盐标记的。β链胰蛋白酶肽段的反相HPLC分析表明，标记仅与II、III和IV中的1 - 缬氨酸残基结合，而V和VI中的1 - 缬氨酸和82 - 赖氨酸均被标记。同样，检测到IV和VI的α链1 - 缬氨酸残基的修饰。组分II和III具有相同的分子式。有证据表明它们是可相互转化的，且它们对应于PLP2 - Hb物种，组分V是贝内施的PLP4 - Hb[《生物化学杂志》，257(1983)1320及其中引用的参考文献]。组分IV是III，每条α链多一个PLP，类似地，VI是V，其α链为单标记。(摘要截断于250字)
