Slaughter R S, Welton A F, Morgan D W
Allergy and Inflammation Research, Hoffmann-La Roche Inc., Nutley, NJ.
Biochim Biophys Acta. 1987 Nov 2;904(1):92-104. doi: 10.1016/0005-2736(87)90090-3.
Sarcolemmal vesicles prepared by a new procedure from bovine tracheal smooth muscle were found to have a Na-Ca exchange activity that is significantly higher than that reported for different preparations from other types of smooth muscle. The exchange process system co-purified with 5'-nucleotidase, a plasma membrane marker enzyme, and was significantly enriched (over 100-fold) compared to mitochondria (cytochrome-c oxidase) but only slightly enriched (4-fold) compared to sarcoplasmic reticulum (NADPH-cytochrome-c reductase). The Na+ dependence of Ca2+ transport was demonstrated through both uptake and efflux procedures. The uptake profile with respect to Ca2+ was monotonic with a linear vo VS. vo.S-1 plot. The resultant Km of Ca2+ from the airway sarcolemmal vesicles (20 microM) was similar in magnitude to the Km of cardiac sarcolemmal vesicles (30 microM). Tracheal vesicles demonstrated a Vmax of 0.3-0.5 nmol.mg-1.s-1 which is significantly higher than that reported in preparations from other smooth muscle types. Furthermore, two processes found to stimulate cardiac Na-Ca exchange, pretreatment with either a mixture of dithiothreitol and Fe2+ or with chymotrypsin, were ineffective on the tracheal smooth muscle. Thus, the Na-Ca exchanger identified in tracheal smooth muscle appears to be different from that observed in cardiac muscle, implying that regulation of this activity may also be different.
通过一种新方法从牛气管平滑肌制备的肌膜囊泡被发现具有钠钙交换活性,该活性显著高于从其他类型平滑肌的不同制剂中报道的活性。该交换过程系统与质膜标记酶5'-核苷酸酶共纯化,与线粒体(细胞色素c氧化酶)相比显著富集(超过100倍),但与肌浆网(NADPH-细胞色素c还原酶)相比仅略有富集(4倍)。通过摄取和流出程序都证明了Ca2+转运对Na+的依赖性。关于Ca2+的摄取曲线是单调的,vo与vo.S-1呈线性关系。气道肌膜囊泡中Ca2+的最终Km(20 microM)在大小上与心脏肌膜囊泡的Km(30 microM)相似。气管囊泡的Vmax为0.3-0.5 nmol.mg-1.s-1,这显著高于从其他平滑肌类型的制剂中报道的值。此外,发现刺激心脏钠钙交换的两个过程,即用二硫苏糖醇和Fe2+的混合物预处理或用胰凝乳蛋白酶预处理,对气管平滑肌无效。因此,在气管平滑肌中鉴定出的钠钙交换器似乎与在心肌中观察到的不同,这意味着这种活性的调节也可能不同。