Hostomský Z, Smrt J, Paces V
Gene. 1985;39(2-3):269-74. doi: 10.1016/0378-1119(85)90322-1.
Enkephalins are pentapeptides with opioid activity that have been found in brain and other neural tissues. They are released by proteolytic processing of the proenkephalin, which contains several enkephalin sequences each flanked by pairs of basic amino acid (aa) residues. We have constructed an artificial variant of the proenkephalin gene by concatenation of synthetic oligodeoxynucleotides (oligo) coding for Met-enkephalin preceded by two arginines. One of the resulting structures, containing eleven enkephalin sequences separated by pairs of arginine codons, was cloned in the expression vector pRE31. The biological activity of enkephalin was detected after the digestion of the isolated plasmid-coded protein with trypsin and carboxypeptidase B. The product of the synthetic gene may thus serve as a defined simplified substrate for the study of the not yet fully understood enzymatic mechanisms of proenkephalin processing.
脑啡肽是具有阿片样活性的五肽，已在脑和其他神经组织中发现。它们由前脑啡肽经蛋白水解加工释放，前脑啡肽包含几个脑啡肽序列，每个序列两侧都有一对碱性氨基酸残基。我们通过连接编码甲硫氨酸脑啡肽且前面有两个精氨酸的合成寡脱氧核苷酸构建了前脑啡肽基因的人工变体。其中一个产生的结构包含由精氨酸密码子对隔开的11个脑啡肽序列，被克隆到表达载体pRE31中。用胰蛋白酶和羧肽酶B消化分离的质粒编码蛋白后，检测到了脑啡肽的生物活性。因此，合成基因的产物可作为一种明确的简化底物，用于研究尚未完全理解的前脑啡肽加工的酶促机制。
