Wuytack F, Kanmura Y, Eggermont J A, Raeymaekers L, Verbist J, Hartweg D, Gietzen K, Casteels R
Laboratorium voor Fysiologie, Katholieke Universiteit Leuven, Belgium.
Biochem J. 1989 Jan 1;257(1):117-23. doi: 10.1042/bj2570117.
Smooth muscle expresses in its endoplasmic reticulum an isoform of the Ca2+-transport ATPase that is very similar to or identical with that of the cardiac-muscle/slow-twitch skeletal-muscle form. However, this enzyme differs from that found in fast-twitch skeletal muscle. This conclusion is based on two independent sets of observations, namely immunological observations and phosphorylation experiments. Immunoblot experiments show that two different antibody preparations against the Ca2+-transport ATPase of cardiac-muscle sarcoplasmic reticulum also recognize the endoplasmic-reticulum/sarcoplasmic-reticulum enzyme of the smooth muscle and the slow-twitch skeletal muscle whereas they bind very weakly or not at all to the sarcoplasmic-reticulum Ca2+-transport ATPase of the fast-twitch skeletal muscle. Conversely antibodies directed against the fast-twitch skeletal-muscle isoform of the sarcoplasmic-reticulum Ca2+-transport ATPase do not bind to the cardiac-muscle, smooth-muscle or slow-twitch skeletal-muscle enzymes. The phosphorylated tryptic fragments A and A1 of the sarcoplasmic-reticulum Ca2+-transport ATPases have the same apparent Mr values in cardiac muscle, slow-twitch skeletal muscle and smooth muscle, whereas the corresponding fragments in fast-twitch skeletal muscle have lower apparent Mr values. This analytical procedure is a new and easy technique for discrimination between the isoforms of endoplasmic-reticulum/sarcoplasmic-reticulum Ca2+-transport ATPases.
平滑肌在内质网中表达一种钙转运ATP酶同工型,它与心肌/慢肌型骨骼肌的钙转运ATP酶同工型非常相似或相同。然而,这种酶与快肌型骨骼肌中的酶不同。这一结论基于两组独立的观察结果,即免疫学观察和磷酸化实验。免疫印迹实验表明,两种针对心肌肌浆网钙转运ATP酶的不同抗体制剂也能识别平滑肌和慢肌型骨骼肌的内质网/肌浆网酶,而它们与快肌型骨骼肌肌浆网钙转运ATP酶的结合非常弱或根本不结合。相反,针对肌浆网钙转运ATP酶快肌型骨骼肌同工型的抗体不与心肌、平滑肌或慢肌型骨骼肌的酶结合。肌浆网钙转运ATP酶的磷酸化胰蛋白酶片段A和A1在心肌、慢肌型骨骼肌和平滑肌中的表观分子量值相同,而在快肌型骨骼肌中相应片段的表观分子量值较低。这种分析方法是一种区分内质网/肌浆网钙转运ATP酶同工型的新颖且简便的技术。