Fisher S A, Ikebe M
Division of Cardiology, Case Western Reserve University, Cleveland, OH 44106, USA.
Biochem Biophys Res Commun. 1995 Dec 14;217(2):696-703. doi: 10.1006/bbrc.1995.2829.
Using a combination of Northern and Western blotting and RT-PCR, we demonstrate the existence of a high molecular mass MLCK, which is expressed during chicken embryogenesis. It is expressed in developing smooth muscle containing tissues, and is detected at low concentrations in adult tissues. Direct sequencing of the RT-PCR product from embryonic tissue RNA revealed that the embryonic, high molecular mass MLCK is indeed the previously cloned "nonmuscle MLCK". Therefore, the high molecular mass MLCK should be termed embryonic/non-muscle MLCK isoform. Curiously, cultured embryonic gizzard and vascular smooth muscle cells express the lower molecular mass smMLCK protein, albeit at lower levels than in the in vivo tissues.
通过结合Northern印迹法、Western印迹法和逆转录聚合酶链反应(RT-PCR),我们证明了一种高分子量肌球蛋白轻链激酶(MLCK)的存在,它在鸡胚胎发育过程中表达。它在发育中的含平滑肌组织中表达,并在成年组织中以低浓度被检测到。对来自胚胎组织RNA的RT-PCR产物进行直接测序表明,胚胎期的高分子量MLCK确实是先前克隆的“非肌肉MLCK”。因此,高分子量MLCK应被称为胚胎/非肌肉MLCK同工型。奇怪的是,培养的胚胎砂囊和血管平滑肌细胞表达较低分子量的平滑肌MLCK蛋白,尽管其水平低于体内组织。