Bogaert R, Tiller G E, Weis M A, Gruber H E, Rimoin D L, Cohn D H, Eyre D R
Department of Orthopaedics, University of Washington, Seattle 98195.
J Biol Chem. 1992 Nov 5;267(31):22522-6.
The spondyloepiphyseal dysplasia subclassification of bone dysplasias includes achondrogenesis, hypochondrogenesis, and spondyloepiphyseal dysplasia congenita. The phenotypic expression of these disorders ranges from mild to perinatal lethal forms. We report the detection and partial characterization of a defect in type II collagen in a perinatal lethal form of hypochondrogenesis. Electrophoresis in sodium dodecyl sulfate-polyacrylamide of CB peptides (where CB represents cyanogen bromide) from type II collagen of the diseased cartilage showed a doublet band for peptide alpha 1(II)CB10 and evidence for post-translational overmodification of the major peptides (CB8, CB10, and CB11) seen as a retarded electrophoretic mobility. Peptide CB10 was digested by endoproteinase Asp-N; and on reverse-phase high pressure liquid chromatography, fragments of abnormal mobility were noted. Sequence analysis of a unique peptide D12 revealed a single amino acid substitution (Gly-->Glu) at position 853 of the triple helical domain. This was confirmed by sequence analysis of amplified COL2A1 cDNA, which revealed a single nucleotide substitution (GGA-->GAA) in 5 of 10 clones. Electron micrographs of the diseased cartilage showed a sparse extracellular matrix and chondrocytes containing dilated rough endoplasmic reticulum, which suggested impaired assembly and secretion of the mutant protein. This case further documents the molecular basis of the spondyloepiphyseal dysplasia spectrum of chondrodysplasias as mutations in COL2A1.
骨发育不良的脊椎骨骺发育不良亚类包括软骨发育不全、低软骨发育不全和先天性脊椎骨骺发育不良。这些疾病的表型表现从轻度到围产期致死型不等。我们报告了在一例围产期致死型低软骨发育不全中检测到II型胶原蛋白缺陷并对其进行部分特征描述。对患病软骨的II型胶原蛋白的CB肽(其中CB代表溴化氰)进行十二烷基硫酸钠-聚丙烯酰胺电泳,结果显示肽α1(II)CB10出现双峰带,并且主要肽段(CB8、CB10和CB11)存在翻译后过度修饰的证据,表现为电泳迁移率延迟。肽CB10被天冬氨酸内肽酶Asp-N消化;在反相高压液相色谱上,发现了迁移率异常的片段。对一个独特的肽段D12进行序列分析,发现在三螺旋结构域的第853位有一个单氨基酸取代(甘氨酸→谷氨酸)。通过对扩增的COL2A1 cDNA进行序列分析证实了这一点,该分析显示在10个克隆中有5个存在单核苷酸取代(GGA→GAA)。患病软骨的电子显微镜照片显示细胞外基质稀疏,软骨细胞含有扩张的粗面内质网,这表明突变蛋白的组装和分泌受损。该病例进一步证明了软骨发育不全的脊椎骨骺发育不良谱系的分子基础是COL2A1中的突变。