Kirino Y, Shimizu H
J Biochem. 1982 Oct;92(4):1287-96. doi: 10.1093/oxfordjournals.jbchem.a134047.
Uptake and release of Ca2+ in heavy and light fractions of fragmented sarcoplasmic reticulum (FSR) isolated from frog and rabbit skeletal muscle was studied under conditions similar to those employed in skinned muscle fiber experiments, where ATP and Mg2+ concentrations were considered to be physiological and free Ca2+ concentration was kept constant during the Ca2+ uptake and release. Ca2+ level in FSR monotonously approached a steady state level which depended only on the final experimental conditions. Heavy fractions, but not light fractions, exhibited characteristics similar to those of Ca2+-induced Ca2+ release reported in skinned fiber studies: i) the rate and steady state level of Ca2+ uptake increased with increase in free Ca2+ concentration in the reaction medium up to 10(-6) M. With further increase in free Ca2+ concentration, the steady state level of Ca2+ taken up decreased while the Ca2+ uptake rate increased. ii) The steady state Ca2+ level was decreased by caffeine but increased by procaine or ruthenium red. Parallel measurement of Ca2+-ATPase activity clearly showed that these drugs modify the Ca2+ efflux but hardly affect the Ca2+-pump activity. It was concluded that the Ca2+-induced Ca2+ release mechanism was in operation at as low as 10(-6) M free Ca2+ concentration. Treatment of FSR with 0.6 M KCl did not have any significant effect.
在类似于用于完整肌纤维实验的条件下,研究了从青蛙和兔子骨骼肌分离出的破碎肌浆网(FSR)重组分和轻组分中Ca2+的摄取和释放。在这些条件下,ATP和Mg2+浓度被认为是生理浓度,并且在Ca2+摄取和释放过程中游离Ca2+浓度保持恒定。FSR中的Ca2+水平单调地接近一个仅取决于最终实验条件的稳态水平。重组分而非轻组分表现出与完整纤维研究中报道的Ca2+诱导的Ca2+释放相似的特征:i)在反应介质中游离Ca2+浓度增加至10(-6) M之前,Ca2+摄取的速率和稳态水平随其增加而增加。随着游离Ca2+浓度进一步增加,摄取的Ca2+稳态水平降低,而Ca2+摄取速率增加。ii)咖啡因降低稳态Ca2+水平,而普鲁卡因或钌红则增加该水平。Ca2+-ATP酶活性的平行测量清楚地表明,这些药物改变Ca2+外流,但几乎不影响Ca2+泵活性。得出的结论是,在低至10(-6) M的游离Ca2+浓度下,Ca2+诱导的Ca2+释放机制仍在起作用。用0.6 M KCl处理FSR没有任何显著影响。