Potier M C, Chelot E, Pekarsky Y, Gardiner K, Rossier J, Turnell W G
Institut Alfred Fessard, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Genomics. 1995 Oct 10;29(3):562-70. doi: 10.1006/geno.1995.9965.
Myosin light chain kinase (MLCK), a key enzyme in muscle contraction, has been shown by immunohistology to be present in neurons and glia. We describe here the cloning of the cDNA for human MLCK from hippocampus, encoding a protein sequence 95% similar to smooth muscle MLCKs but less than 60% similar to skeletal muscle MLCKs. The cDNA clone detected two RNA transcripts in human frontal and entorhinal cortex, in hippocampus, and in jejunum, one corresponding to MLCK and the other probably to telokin, the carboxy-terminal 154 codons of MLCK expressed as an independent protein in smooth muscle. Levels of expression were lower in brain compared to smooth muscle. We show that within the protein sequence, a motif of 28 or 24 residues is repeated five times, the second repeat ending with the putative methionine start codon. These repeats overlap with a second previously reported module of 12 residues repeated five times in the human sequence. In addition, the acidic C-terminus of all MLCKs from both brain and smooth muscle resembles the C-terminus of tubulins. The chromosomal localization of the gene for human MLCK is shown to be at 3qcen-q21, as determined by PCR and Southern blotting using two somatic cell hybrid panels.
肌球蛋白轻链激酶(MLCK)是肌肉收缩中的一种关键酶,免疫组织学研究表明它存在于神经元和神经胶质细胞中。我们在此描述了从人脑海马体中克隆人MLCK的cDNA,其编码的蛋白质序列与平滑肌MLCK有95%的相似性,但与骨骼肌MLCK的相似性不到60%。该cDNA克隆在人额叶和内嗅皮质、海马体以及空肠中检测到两种RNA转录本,一种对应于MLCK,另一种可能对应于telokin,即MLCK的羧基末端154个密码子在平滑肌中作为一种独立蛋白质表达。与平滑肌相比,脑中的表达水平较低。我们发现,在蛋白质序列中,一个由28个或24个残基组成的基序重复了五次,第二个重复序列以推定的甲硫氨酸起始密码子结束。这些重复序列与之前报道的另一个在人类序列中重复五次的12个残基的模块重叠。此外,来自脑和平滑肌的所有MLCK的酸性C末端类似于微管蛋白的C末端。通过使用两个体细胞杂交板进行PCR和Southern印迹分析,确定人MLCK基因的染色体定位在3qcen-q21。