饱和转移电子顺磁共振实验方法的研究

本文以马来酰亚胺自旋标记的人血红蛋白样品为例,阐述了饱和转移电子顺磁共振实验技术,其中包括样品制备、测量要点和波谱参数的测定。并探讨了实验条件对波谱的影响。     

Mg~(2+)通过磷脂影响脂酶体中H~+-ATP酶的慢运动

本文研究Mg~(2+)对H~+-ATP酶在脂酶体中慢运动的影响。在不同Mg~(2+)浓度及不同温度下,测量了运用保温法嵌入脂酶体中的马来酰亚胺自旋标记的H~+-ATP酶的ST-EPR谱。由ST-EPR谱的诊断参数L″/L,C′/C与Mg~(2+)浓度以及温度的关系,并对照根据相同条件下测得的5NS-EPR谱超精细分裂而得到的序参数S与Mg~(2+)浓度以及温度的关系,分析得出:增加脂酶体中的Mg~(2+)浓度与降低测量温度都导致H~+-ATP酶在膜脂中的运动相关时间增加。进一步分析还得到H~+-ATP酶在膜脂中旋转扩散运动的快慢与周围膜脂的活化能大小有关。在脂酶体中Mg~(2+)浓度的变化将导致膜脂活化能发生改变,从而影响H~+-ATP酶在膜中的慢运动状态。     

Mg~(2+)-MEDIATED CHANGES IN FLUIDITY OF PHOSPHOLIPID DECREASE THE SLOW MOTION OF THE INCORPORATED H~+-ATPase IN LIPOSOMES

The effect of Mg~(2+) on the slow motion of H~+-ATPase in proteoliposome was studied. The ST-ESR spectra of H~+-ATPase labeled with maleimide and incorporated in liposomes at various Ma~(2+)concentrations with incubation method were measured. The relationship between the diagnostic param-eters L"/L, C'/C derived from ST-ESR spectra and Mg~(2+) concentrations or temperatures was com-pared with that of the order parameter S derived from 5NS-ESR spectra under the same conditions. Obtain-ed results clearly showed that the increase of Mg~(2+) concentration or the decrease of temperature maycause the increase of the rotational correlation time of H~+-ATPase in proteoliposome. And the rota-tional diffusion rate of H~+-ATPase in membrane lipid is related to the activation energy of the sur-rounding lipid. The change of Mg~(2+) concentration in proteoliposome leads to a change of activarionenergy and influence in turn the slow motion of H~+-ATPase in 1ipid.