Molecular and immunochemical evidences demonstrate that endooligopeptidase A is the predominant cytosolic oligopeptidase of rabbit brain
Molecular and immunochemical evidences demonstrate that endooligopeptidase A is the predominant cytosolic oligopeptidase of rabbit brain
复制标题
DOI：
10.1006/bbrc.2000.2243
复制
发表时间：
2000-03-05
影响因子：
3.1
通讯作者：
de Camargo, ACM
中科院分区：
文献类型：
--
作者：
Hayashi, MAF;Portaro, FCV;de Camargo, ACM
寡肽酶是一种不攻击蛋白质的组织内肽酶，可能参与多肽激素和神经肽的成熟和降解。兔脑内寡肽酶A和大鼠睾丸可溶性金属肽酶(EC 3.4.24.15)是硫醇激活的寡肽酶，它能从许多阿片肽中产生脑啡肽，并通过降解相同的肽键来灭活缓激肽和神经降压素。用纯化的兔脑内寡肽酶A制备的单特异性抗体能够鉴定出一个2.3kb的编码512个氨基酸的截短酶的cDNA，显示出与内寡肽酶A相同的酶特性。尽管采取了各种策略，但直到现在仍未能克隆出全长的cDNA。推导出的氨基酸序列与大鼠睾丸金属内肽酶序列没有相似之处，但存在典型的金属蛋白酶共有序列[HEXXH]。抗重组内寡肽酶A抗体特异性地抑制其自身活性，使兔脑胞浆中硫醇激活的寡肽酶活性降低到30%以下。内寡肽酶A的组织分布分析表明，该酶主要在中枢神经系统表达，而可溶性金属EC3.4.24.15主要在外周组织表达。(C)2000年学术出版社。
Oligopeptidases are tissue endopeptidases that do not attack proteins and are likely to be involved in the maturation and degradation of peptide hormones and neuropeptides. The rabbit brain endooligopeptidase A and the rat testes soluble metallopeptidase (EC 3.4.24.15) are thiol-activated oligopeptidases which are able to generate enkephalin from a number of opiod peptides and to inactivate bradykinin and neurotensin by hydrolyzing the same peptide bonds. A monospecific antibody raised against the purified rabbit brain endooligopeptidase A allowed the identification of a 2.3 kb cDNA coding for a truncated enzyme of 512 amino acids, displaying the same enzymatic features as endooligopeptidase A. In spite of all efforts, employing several strategies, the full-length cDNA could not be cloned until now. The analysis of the deduced amino acid sequence showed no similarity to the rat testes metalloendopeptidase sequence, except for the presence of the typical metalloprotease consensus sequence [HEXXH]. The antibody raised against recombinant endooligopeptidase A specifically inhibited its own activity and reduced the thiol-activated oligopeptidase activity of rabbit brain cytosol to less than 30%. Analysis of the endooligopeptidase A tissue distribution indicated that this enzyme is mainly expressed in the CNS, whereas the soluble metallo EC 3.4.24.15 is mainly expressed in peripheral tissues. (C) 2000 Academic Press.