Ishii Y, Lehrer S S
Department of Muscle Research, Boston Biomedical Research Institute, Massachusetts 02114.
Biophys J. 1989 Jul;56(1):107-14. doi: 10.1016/S0006-3495(89)82655-4.
Tropomyosin (Tm) paracrystal formation induced by Mg2+ was studied by monitoring increases in light scattering. Paracrystals formed above a critical Tm concentration with lag phases in the time courses at pH 7.5 and 6.0, indicating that condensation polymerization processes are involved. The kinetic data at pH 7.5 reasonably fit a model in which nucleation and elongation are taken into account. The rate and extent of light scattering increased at low [Mg2+] and decreased at high [Mg2+] with a maximum at [Mg2+] = 15 mM, indicating different effects of Mg2+ in the two [Mg2+] ranges. The paracrystals were destabilized by increasing the salt concentration and decreasing the temperature. Mg2+ produces paracrystals at pH 6.0 and pH 7.5 by different kinetic mechanisms. Different Tm intermolecular interactions at the two pH values were indicated by studies of the excimer fluorescence of pyrene-labeled Tm and by effects of salt and temperature on the kinetics. At pH 6.0 Tm more readily formed paracrystals with decreased electrostatic effects. Effects of troponin on Mg2+-paracrystal formation of Tm at the two pH values correlated with the known differences in paracrystal structure when troponin is bound to Tm.
通过监测光散射的增加来研究镁离子诱导的原肌球蛋白(Tm)副晶形成。在pH 7.5和6.0时,副晶在临界Tm浓度以上形成,且时间进程中有滞后阶段,表明涉及缩合聚合过程。pH 7.5时的动力学数据合理地符合一个考虑了成核和延伸的模型。在低镁离子浓度下,光散射的速率和程度增加,在高镁离子浓度下降低,在镁离子浓度为15 mM时达到最大值,表明镁离子在这两个镁离子浓度范围内有不同的作用。通过增加盐浓度和降低温度,副晶会变得不稳定。镁离子在pH 6.0和pH 7.5时通过不同的动力学机制产生副晶。芘标记的Tm的准分子荧光研究以及盐和温度对动力学的影响表明,在这两个pH值下Tm分子间相互作用不同。在pH 6.0时,Tm更容易形成副晶,静电效应降低。肌钙蛋白在这两个pH值下对Tm的镁离子副晶形成的影响与肌钙蛋白与Tm结合时副晶结构的已知差异相关。