Lysyl oxidase-like protein from bovine aorta - Isolation and maturation to an active form by bone morphogenetic protein-1
Lysyl oxidase-like protein from bovine aorta - Isolation and maturation to an active form by bone morphogenetic protein-1
复制标题
DOI：
10.1074/jbc.m109499200
复制
发表时间：
2001-12-28
影响因子：
4.8
通讯作者：
Font, B
中科院分区：
文献类型：
--
作者：
Borel, A;Eichenberger, D;Font, B
最近已经描述了几种编码赖氨酰氧化酶样蛋白的cDNA。它们推导的氨基酸序列的特征是C-末端区域（对应于赖氨酰氧化酶家族催化结构域）具有很强的相似性，而N-末端区域则存在显着差异。赖氨酰氧化酶除了具有传统的交联作用外，还具有不同的生物学功能。为了回答这些不同的功能是否由不同的赖氨酰氧化酶执行的问题，需要这些酶的纯化和活性形式。目前，只有经典形式的赖氨酰氧化酶已被纯化和表征。本研究的目的是分离和表征赖氨酰氧化酶样蛋白。鉴于与其他赖氨酰氧化酶的C-末端结构域的强序列同源性，我们选择使用对酶原的N-末端结构域具有特异性的抗体从牛主动脉纯化蛋白质。我们已经分离出一种56 kDa的蛋白质，通过氨基酸测序鉴定为牛赖氨酰氧化酶样前体，其在位置89-91处的Arg-Arg-Arg序列处被弗林蛋白酶样活性裂解，如在N-末端残基去封闭后所揭示的。免疫纯化的蛋白在很大程度上是无活性的，但在体外进一步处理的骨形态发生蛋白-1导致的酶，是活跃的弹性蛋白和胶原底物。
Recently several cDNAs have been described encoding lysyl oxidase-like proteins. Their deduced amino acid sequences are characterized by a strong similarity in the C-terminal region, corresponding to the lysyl oxidase family catalytic domain, and by marked differences in the N-terminal regions. Different biological functions have been described for lysyl oxidases in addition to their traditionally assumed cross-linking role. To answer the question of whether these different functions are carried out by different lysyl oxidases, purified and active forms of these enzymes are required. At present only the classical form of lysyl oxidase has been purified and characterized. The purpose of this study was to isolate and characterize the lysyl oxidase-like protein. In view of the strong sequence homology with the C-terminal domain of other lysyl oxidases, we chose to purify the protein from bovine aorta using antibodies specific to the N-terminal domain of the proenzyme. We have isolated a 56-kDa protein identified by amino acid sequencing as the bovine lysyl oxidase-like precursor, which is cleaved at the Arg-Arg-Arg sequence at positions 89-91 by a furin-like activity, as revealed after deblocking of the N-terminal residue. The immunopurified protein was largely inactive, but further processing in vitro by bone morphogenetic protein-1 led to an enzyme that was active on elastin and collagen substrates.