Characterization of Endothelin-converting Enzyme-2
Characterization of Endothelin-converting Enzyme-2
复制标题
内皮素转换酶 2 的表征
DOI：
--
复制
发表时间：
2003
影响因子：
4.8
通讯作者：
L. Devi
中科院分区：
文献类型：
--
作者：
N. Mzhavia;Hui Pan;F. Che;L. Fricker;L. Devi
大多数神经内分泌肽是由碱性残基裂解位点的前体蛋白分解而产生的。属于枯草杆菌丝氨酸蛋白酶家族的前激素转换酶主要负责这些“经典部位”的加工。除了经典的裂解作用外，生物活性多肽的一个子集是通过在“非经典”位点进行加工而产生的。负责这些分裂的蛋白水解酶还没有被很好地研究。一些金属蛋白水解酶家族的成员被认为与非经典加工有关。其中，内皮素转换酶-2(ECE2)是一个很好的候选者，因为它具有神经内分泌分布和酸性最适pH。为了研究该酶在神经肽加工中的作用，我们对重组酶进行了纯化，并对其催化活性进行了鉴定。在酸性pH条件下，通过Trp21和Val22之间的裂解，纯化的ECE2能有效地将大的内皮素-1转化为内皮素-1。为了鉴定ECEs-2的底物专一性，我们使用了一组42个多肽作为底物的质谱仪来鉴定产物。在这42个多肽中，只有10个被欧洲经委会-2处理过。对切割位点周围残基的比较表明，ECE2显示出独特的切割位点选择性，这与ECE1的选择性有关，但又不同。ECE-2耐受P1-位的多种氨基酸，而偏爱P1‘-位的脂肪族/芳香族残基。然而，只有一小部分含脂肪/芳香氨基酸的位点被切割，这表明在P1-和P1‘-位置之外还有额外的限制。该酶能够从多肽中间体中产生许多生物活性多肽，这表明该酶在调节多肽的生物合成中起着重要的作用。此外，ECA-2处理前脑啡肽衍生的牛肾上腺髓质多肽，这种处理导致已知具有不同受体选择性的多肽产品。最后，欧洲经委会-2将PEN-LEN(前激素转换酶1的内源性抑制物)加工成不抑制该酶的产物。综上所述，这些结果与ECA-2在处理非经典位点的调节肽过程中的一个重要角色是一致的。
Most neuroendocrine peptides are generated by proteolysis of the precursors at basic residue cleavage sites. Prohormone convertases belonging to the subtilisin family of serine proteases are primarily responsible for processing at these “classical sites.” In addition to the classical cleavages, a subset of bioactive peptides is generated by processing at “nonclassical” sites. The proteases responsible for these cleavages have not been well explored. Members of several metalloprotease families have been proposed to be involved in nonclassical processing. Among them, endothelin-converting enzyme-2 (ECE-2) is a good candidate because it exhibits a neuroendocrine distribution and an acidic pH optimum. To examine the involvement of this protease in neuropeptide processing, we purified the recombinant enzyme and characterized its catalytic activity. Purified ECE-2 efficiently processes big endothelin-1 to endothelin-1 by cleavage between Trp21 and Val22 at acidic pH. To characterize the substrate specificity of ECE-2, we used mass spectrometry with a panel of 42 peptides as substrates to identify the products. Only 10 of these 42 peptides were processed by ECE-2. A comparison of residues around the cleavage site revealed that ECE-2 exhibits a unique cleavage site selectivity that is related to but distinct from that of ECE-1. ECE-2 tolerates a wide range of amino acids in the P1-position and prefers aliphatic/aromatic residues in the P1′-position. However, only a small fraction of the aliphatic/aromatic amino acid-containing sites were cleaved, indicating that there are additional constraints beyond the P1- and P1′-positions. The enzyme is able to generate a number of biologically active peptides from peptide intermediates, suggesting an important role for this enzyme in the biosynthesis of regulatory peptides. Also, ECE-2 processes proenkephalin-derived bovine adrenal medulla peptides, and this processing leads to peptide products known to have differential receptor selectivity. Finally, ECE-2 processes PEN-LEN, an endogenous inhibitor of prohormone convertase 1, into products that do not inhibit the enzyme. Taken together, these results are consistent with an important role for ECE-2 in the processing of regulatory peptides at nonclassical sites.
影响因子：
4.1
作者：
Mzhavia, N;Qian, Y;Fricker, LD
通讯作者：
Fricker, LD
DOI：
--
发表时间：
1991-04
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
B. Eipper;S. Perkins;E. Husten;R. C. Johnson;H. Keutmann;R. Mains
通讯作者：
B. Eipper;S. Perkins;E. Husten;R. C. Johnson;H. Keutmann;R. Mains
人金属蛋白酶 1 的克隆、表达和表征：金属内蛋白酶 Pitrilysin 家族的新成员。
DOI：
10.1089/104454999315268
发表时间：
1999
期刊：
DNA and cell biology.
影响因子：
--
作者：
Mzhavia,N;Berman,YL;Qian,Y;Yan,L;Devi,LA
通讯作者：
Devi,LA