THE PRIMARY STRUCTURE OF HUMAN DOPAMINE-BETA-HYDROXYLASE - INSIGHTS INTO THE RELATIONSHIP BETWEEN THE SOLUBLE AND THE MEMBRANE-BOUND FORMS OF THE ENZYME
THE PRIMARY STRUCTURE OF HUMAN DOPAMINE-BETA-HYDROXYLASE - INSIGHTS INTO THE RELATIONSHIP BETWEEN THE SOLUBLE AND THE MEMBRANE-BOUND FORMS OF THE ENZYME
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DOI：
10.1002/j.1460-2075.1987.tb02734.x
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发表时间：
1987-12-01
期刊：
影响因子：
11.4
通讯作者：
MALLET, J
中科院分区：
文献类型：
--
作者：
LAMOUROUX, A;VIGNY, A;MALLET, J
一个完整的多巴胺- β。从人嗜铬细胞瘤中分离到-羟化酶（DBH） cDNA克隆。gt11图书馆。结构和功能证据都证实了克隆的真实性：(1)阳性克隆产生的融合蛋白选择抗体沉淀DBH活性，(2)三个内部DBH色氨酸的序列包含在推断的DBH序列中，(3)先前报道的牛DBH n端15个氨基酸显示与预测的人类DBH几乎完全一致。DBH的多肽链包括578个氨基酸，对应于一个64 862道尔顿的未修饰蛋白，前面是一个具有25个残基的裂解信号肽。DBH以膜结合和可溶性两种形式存在。亲水图显示除信号肽外无明显疏水段。S1图谱分析显示，DBH mRNA的5‘和3’端没有多样性。结合现有的生化数据，这些观察结果表明，DBH的膜附着可能是翻译后修饰的结果，字形化是最有可能的候选人。比较氨基酸序列分析表明，DBH与其他儿茶酚胺合成酶、酪氨酸羟化酶和苯乙醇胺- n -甲基转移酶没有同源性。
A full length dopamine-.beta.-hydroxylase (DBH) cDNA clone was isolated from a human pheochromocytoma .lambda.gt11 library. Both structural and functional evidence confirms the authenticity of the clone: (i) antibodies selected with fusion proteins generated by positive clones precipitate DBH activity, (ii) the sequence of three internal DBH tryptic peptides are included in the deduced DBH sequence, (iii) the previously reported N-terminal 15 amino acids of bovine DBH exhibits a nearly complete identity with that predicted for human DBH. The polypeptide chain of DBH comprises 578 amino acids corresponding to an unmodified protein of 64 862 daltons and is preceded by a cleaved signal peptide of 25 residues. DBH exists in both membrane-bound and soluble forms. The hydropathy plot reveals no obvious hydrophobic segment, except the signal peptide. S1 mapping analysis indicates no diversity in the 5'' and 3'' extremities of the DBH mRNA. Taken together with available biochemical data, these observations suggest that the membrane attachment of DBH probably results from a post-translational modification, glypiation being the most likely candidate. Comparative amino acid sequence analysis establishes that DBH shares no homology with the other catecholamine synthesizing enzymes, tyrosine hydroxylase and phenylethanolamine-N-methyl transferase.