van Ginkel L A, Somers H H, Loeber J G
National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
Acta Endocrinol (Copenh). 1991 Jul;125(1):73-9. doi: 10.1530/acta.0.1250073.
The heterogeneity of human luteinizing hormone was investigated with high performance hydrophobic interaction chromatography on a TSK phenyl-5PW column using an ammonium sulphate gradient. Recovery of individual subunits, expressed as immunochemical activity, was in the order of 90%. Recovery of the intact hormone was less, approximately 25%. The technique appeared to be independent of the charge heterogeneity of the individual subunits. Each component as obtained by hydrophobic interaction chromatography showed considerable charge heterogeneity when studied by subsequent isoelectric focusing. Nonetheless, all chromatography fractions derived from either subunit showed the same collection of isoelectric values, quantitatively only differences for the beta-subunits could be detected. Heterogeneity of the individual subunits was clearly demonstrated after incubation of the preparations at 37 or 56 degrees C. The heterogeneity of the beta-subunit observed after incubation at 56 degrees C was different to the heterogeneity after incubation at 37 degrees C. After incubation at 56 degrees C, an additional component, with a longer retention time, developed directly from the intact molecule. The component is unstable and is transformed to one of the components detected also after incubation at 37 degrees C. Based on these results the existence of two populations of intact LH molecules with respect to their thermal stability is hypothesized.
采用硫酸铵梯度，在TSK苯基-5PW柱上通过高效疏水相互作用色谱法研究了人促黄体生成素的异质性。以免疫化学活性表示的各个亚基的回收率约为90%。完整激素的回收率较低，约为25%。该技术似乎与各个亚基的电荷异质性无关。通过疏水相互作用色谱法获得的每个组分在随后的等电聚焦研究中显示出相当大的电荷异质性。尽管如此，来自任一亚基的所有色谱馏分都显示出相同的等电值集合，仅在定量上可检测到β亚基的差异。在37或56℃孵育制剂后，清楚地证明了各个亚基的异质性。在56℃孵育后观察到的β亚基的异质性与在37℃孵育后的异质性不同。在56℃孵育后，一个保留时间更长的额外组分直接从完整分子中产生。该组分不稳定，会转化为在37℃孵育后也检测到的组分之一。基于这些结果，推测存在两种关于其热稳定性的完整促黄体生成素分子群体。
