Thulborn K R, Minasian E, Leach S J
Biochim Biophys Acta. 1979 Jun 19;578(2):476-83. doi: 10.1016/0005-2795(79)90177-6.
Leghaemoglobin from the subclover, Trifolium subterraneum cultivar Woogenellup, has been fractionated into at least four electrophoretically distinct components using the ion-exchange chromatographic procedure described by Appleby et al. (Appebly, C.A., Nicola, N.A., Hurrell, J.G.R. and Leach, S.J. (1975) Biochemistry 14, 4444--4450) for soybean leghaemoglobins. Unlike those of soybean, the subclover leghaemoglobins showed no evidence of autoxidation under identical isolation procedures, implying that these proteins have an unusually stable ferrous oxidation state. Circular dichroism in the far-ultraviolet (200--240 nm) indicated a high helicity (approx. 70%) as has been reported for other species of leghaemoglobins. However, circular dichroism in the near-ultraviolet region (240--300 nm) indicated that the haem-protein interactions may be considerably different in the subclover leghaemoglobins and this may explain their atypical resistance to autoxidation and the absence of nicotinate binding in these proteins.
利用Appleby等人（Appleby, C.A., Nicola, N.A., Hurrell, J.G.R.和Leach, S.J.（1975年）《生物化学》14卷，4444 - 4450页）描述的用于大豆豆血红蛋白的离子交换色谱法，已将来自地下三叶草（Trifolium subterraneum）品种Woogenellup的豆血红蛋白分离成至少四种电泳性质不同的组分。与大豆豆血红蛋白不同，在相同的分离程序下，地下三叶草豆血红蛋白没有自氧化的迹象，这意味着这些蛋白质具有异常稳定的亚铁氧化态。远紫外区（200 - 240纳米）的圆二色性表明其具有高螺旋度（约70%），这与其他种类的豆血红蛋白的情况一致。然而，近紫外区（240 -
