Department of Biological Sciences, Biomolecular Research Center and Musculoskeletal Research Institute, Boise State University, Boise, ID 83725-1515, USA.
Proteomics. 2011 Dec;11(24):4660-76. doi: 10.1002/pmic.201100058. Epub 2011 Nov 23.
Cartilage plays an essential role during skeletal development within the growth plate and in articular joint function. Interactions between the collagen fibrils and other extracellular matrix molecules maintain structural integrity of cartilage, orchestrate complex dynamic events during embryonic development, and help to regulate fibrillogenesis. To increase our understanding of these events, affinity chromatography and liquid chromatography/tandem mass spectrometry were used to identify proteins that interact with the collagen fibril surface via the amino terminal domain of collagen α1(XI) a protein domain that is displayed at the surface of heterotypic collagen fibrils of cartilage. Proteins extracted from fetal bovine cartilage using homogenization in high ionic strength buffer were selected based on affinity for the amino terminal noncollagenous domain of collagen α1(XI). MS was used to determine the amino acid sequence of tryptic fragments for protein identification. Extracellular matrix molecules and cellular proteins that were identified as interacting with the amino terminal domain of collagen α1(XI) directly or indirectly, included proteoglycans, collagens, and matricellular molecules, some of which also play a role in fibrillogenesis, while others are known to function in the maintenance of tissue integrity. Characterization of these molecular interactions will provide a more thorough understanding of how the extracellular matrix molecules of cartilage interact and what role collagen XI plays in the process of fibrillogenesis and maintenance of tissue integrity. Such information will aid tissue engineering and cartilage regeneration efforts to treat cartilage tissue damage and degeneration.
软骨在生长板的骨骼发育和关节功能中起着至关重要的作用。胶原纤维和其他细胞外基质分子之间的相互作用维持着软骨的结构完整性，协调胚胎发育过程中的复杂动态事件，并有助于调节原纤维形成。为了增加对这些事件的理解，使用亲和层析和液相色谱/串联质谱法来鉴定与通过胶原α 1(XI)的氨基末端结构域相互作用的蛋白质，该蛋白结构域在软骨的异质胶原纤维表面上显示。使用高离子强度缓冲液在匀浆中从胎牛软骨中提取蛋白质，根据对胶原α 1(XI)的氨基末端非胶原结构域的亲和力进行选择。MS 用于确定用于蛋白质鉴定的胰蛋白酶片段的氨基酸序列。鉴定为与胶原α 1(XI)的氨基末端结构域直接或间接相互作用的细胞外基质分子和细胞蛋白，包括蛋白聚糖、胶原和基质细胞分子，其中一些也在原纤维形成中发挥作用，而其他分子则已知在组织完整性的维持中发挥作用。这些分子相互作用的特征将提供更深入的了解，即软骨的细胞外基质分子如何相互作用以及胶原 XI 在原纤维形成和组织完整性维持过程中发挥什么作用。这些信息将有助于组织工程和软骨再生努力，以治疗软骨组织损伤和退化。
