Post-translational modification of bone morphogenetic protein-1 is required for secretion and stability of the protein
Post-translational modification of bone morphogenetic protein-1 is required for secretion and stability of the protein
复制标题
DOI：
10.1074/jbc.m207342200
复制
发表时间：
2002-11-08
影响因子：
4.8
通讯作者：
Kadler, KE
中科院分区：
文献类型：
--
作者：
Garrigue-Antar, L;Hartigan, N;Kadler, KE
骨形态发生蛋白（BMP）-1是一种糖基化的金属蛋白酶，它是合成正常细胞外基质的基础，因为它切割I型前胶原以及其他前体蛋白。序列分析表明BMP-1具有6个潜在的N-连接糖基化位点（即NXS/T），即：Asn（91）（前结构域）、Asn（142）（金属蛋白酶结构域）、Asn（332）和Asn（363）（CUB 1结构域）、Asn（599）（CUB 3结构域）和C-末端特异性结构域中的Asn（726）。在这项研究中，我们发现，所有这些网站是N-糖基化的复杂型寡糖含有唾液酸，除了Asn（726），大概是因为脯氨酸发生在C-末端的苏氨酸的共识序列。缺乏所有糖基化位点或三个CUB特异性位点的重组BMP-1分子不分泌。缺乏CUB糖基化的BMP-1易位到蛋白酶体中进行降解。缺乏个别糖基化位点的BMP-1分子被有效地分泌，并表现出完整的前胶原C-蛋白酶活性，但N332 Q和N599 Q表现出较慢的切割速率。缺乏任何一个CUB特异性糖基化位点的BMP-1分子对热变性敏感。研究表明，BMP-1的CUB结构域中的糖基化位点对于该分子的分泌和稳定性是重要的。
Bone morphogenetic protein (BMP)-1 is a glycosylated metalloproteinase that is fundamental to the synthesis of a normal extracellular matrix because it cleaves type I procollagen, as well as other precursor proteins. Sequence analysis suggests that BMP-1 has six potential N-linked glycosylation sites (i.e. NXS/T) namely: Asn(91) (prodomain), Asn(142) (metalloproteinase domain), Asn(332) and Asn(363) (CUB1 domain), Asn(599) (CUB3 domain), and Asn(726) in the C-terminal-specific domain. In this study we showed that all these sites are N-glycosylated with complex-type oligosaccharides containing sialic acid, except Asn(726) presumably because proline occurs immediately C-terminal of threonine in the consensus sequence. Recombinant BMP-1 molecules lacking all glycosylation sites or the three CUB-specific sites were not secreted. BMP-1 lacking CUB glycosylation was translocated to the proteasome for degradation. BMP-1 molecules lacking individual glycosylation sites were efficiently secreted and exhibited full procollagen C-proteinase activity, but N332Q and N599Q exhibited a slower rate of cleavage. BMP-1 molecules lacking any one of the CUB-specific glycosylation sites were sensitive to thermal denaturation. The study showed that the glycosylation sites in the CUB domains of BMP-1 are important for secretion and stability of the molecule.