GROWTH AND DIFFERENTIATION FACTORS IN BONE AND CARTILAGE

该项目的目标是调查增长的作用， 骨和软骨中的分化因子。 目前项目 调查的主要结果是：1）隔离， 从骨基质中纯化成骨蛋白。 成骨素一直是 从牛、猪和人骨基质中分离和纯化。 通过在肝素琼脂糖凝胶上的亲和层析纯化成骨素， 羟基磷灰石和Sephacryl S-200。 牛成骨素进一步 通过制备型SDS-凝胶电泳纯化。 成骨素活性为 位于30和40 kDa之间的区域。 确定胰蛋白酶肽的数目。 目前的工作是针对 评估结构-功能关系; 2）生产和 合成肽的抗体的表征。 氨基和羧基 已经合成了末端肽和内部肽， 引出的 这些抗血清正在由一个 3）成骨细胞的克隆和表达， 重组DNA方法。 已成功克隆了骨异黄酮， 目前正在尝试以瞬时和稳定的方式表达 4）成骨素（一种细胞外骨）的亲和力 启动骨分化的基质相关蛋白，用于凝集素。 部分纯化的成骨蛋白与伴刀豆球蛋白A结合但不与小麦结合 胚芽凝集素或大豆凝集素。 成骨素是一种糖蛋白， 伴刀豆球蛋白A琼脂糖色谱法是一种有用的技术; 5） 肝素亲和法纯化大鼠牙基质成骨蛋白 6）刺激成骨细胞的表达， 通过成骨素体外软骨形成表型，和7）鸡的反应 肢芽中胚层细胞转化为转化生长因子β 1型。 这项调查的意义在于纠正 骨科颅面畸形、口腔手术和骨折修复 手术

The objective of the project is to investigate the role of growth and differentiation factors in bone and cartilage. The projects currently under investigation with the salient findings are: 1) Isolation and purification of osteogenins from bone matrix. Osteogenins have been isolated and purified from bovine, porcine and human bone matrix. Osteogenin was purified by affinity chromatography on heparin Sepharose, hydroxyapatite and Sephacryl S-200. Bovine osteogenin was further purified by preparative SDS-gel electrophoresis. Osteogenin activity was localized in a zone between 30 and 40 k Da. Amino acid sequence of a number of tryptic peptides were determined. Current work is directed towards assessing structure-function relationships; 2) Production and characterization of antibodies to synthetic peptides. Amino and carboxy terminal and internal peptides have been synthesized and antisera elicited. These antisera are being systematically scrutinized by a battery of techniques; 3) Cloning and expression of osteogenis by recombinant DNA methods. Ostoegenin has been successfully cloned and currently attempts are being made to express in transient and stable expression systems; 4) Affinity of osteogenin, an extracellular bone matrix associated protein initiating bone differentiation, for lectins. Partially purified osteogenin binds to concanavalin A but not to wheat germ agglutins or soybean lectin. Osteogenin is a glycoprotein and concanavalin A Sepharose chromatography is a useful technique; 5) Purification of osteogenin form rat tooth matrix by heparin affinity chromatography; 6) Stimulation of the expression of osteogenic and chondrogenic phenotypes in vitro by osteogenin, and 7) Response of chick limb bud mesodermal cells to transforming growth factor beta-type 1. The significance of this investigation is in the realms of correction of craniofacial anomalies, oral surgery and fracture repair in orthopaedic surgery.