Sercu S, Lambeir A M, Steenackers E, El Ghalbzouri A, Geentjens K, Sasaki T, Oyama N, Merregaert J
Laboratory of Molecular Biotechnology, Department of Biomedical Sciences, University of Antwerp, Wilrijk, Belgium.
Matrix Biol. 2009 Apr;28(3):160-9. doi: 10.1016/j.matbio.2009.02.003. Epub 2009 Mar 9.
The extracellular matrix protein 1 (ECM1) is an 85 kDa secreted glycoprotein, comprising four variants and playing a pivotal role in endochondral bone formation, angiogenesis, and tumour biology. A computational model for the three-dimensional structure of ECM1a was determined to identify the potential and/or concealed region(s) for binding with candidate partners in human skin. Multiple alignments for the secondary structure of ECM1a and b revealed similarity with serum albumin. The N-terminal domain of ECM1a consists mainly of alpha-helices (alphaD1), while the remaining three domains, namely serum albumin subdomain-like (SASDL) domains 2-4, were topologically comparable with the subdomain of the third serum albumin domain. Yeast-two-hybrid screening of a human foreskin cDNA library using both full-length ECM1a and the hot spot region for ECM1 gene mutations in lipoid proteinosis, an autosomal recessive genodermatosis (complete SASDL2 and the linker to SASDL3: aa177-aa361), as bait, isolated seven extracellular proteins. The site-specific interaction of ECM1a with two of these candidate binders, laminin 332 beta-3 chain and fibulin-3, was confirmed by in vitro and in vivo co-immunoprecipitation experiments. Immunohistologically both binders co-localized with ECM1 in human skin. Together, ECM1 is a multifunctional binding core and/or a scaffolding protein interacting with a variety of extracellular and structural proteins, contributing to the maintenance of skin integrity and homeostasis. Hence, disruption of the ECM1 function may cause the failure of multi-communication among the surrounding skin interstitial molecules, as seen in lipoid proteinosis pathology.
细胞外基质蛋白1(ECM1)是一种85 kDa的分泌型糖蛋白,由四种变体组成,在软骨内骨形成、血管生成和肿瘤生物学中起关键作用。确定了ECM1a三维结构的计算模型,以识别其在人皮肤中与候选配体结合的潜在和/或隐蔽区域。ECM1a和b二级结构的多重比对显示与血清白蛋白相似。ECM1a的N端结构域主要由α螺旋(αD1)组成,而其余三个结构域,即血清白蛋白亚结构域样(SASDL)结构域2-4,在拓扑结构上与第三血清白蛋白结构域的亚结构域相当。使用全长ECM1a和类脂蛋白沉积症(一种常染色体隐性遗传性皮肤病)中ECM1基因突变的热点区域(完整的SASDL2和与SASDL3的连接区:第177位氨基酸至第361位氨基酸)作为诱饵,对人包皮cDNA文库进行酵母双杂交筛选,分离出七种细胞外蛋白。通过体外和体内共免疫沉淀实验证实了ECM1a与其中两种候选结合蛋白(层粘连蛋白332β-3链和纤连蛋白-3)的位点特异性相互作用。免疫组织化学显示,这两种结合蛋白在人皮肤中均与ECM1共定位。总之,ECM1是一种多功能结合核心和/或支架蛋白,与多种细胞外和结构蛋白相互作用,有助于维持皮肤的完整性和内环境稳定。因此,ECM1功能的破坏可能导致周围皮肤间质分子之间多重通讯的失败,如在类脂蛋白沉积症病理学中所见。