牛鼻软骨蛋白聚糖的电泳异质性

The electrophoretic heterogeneity of bovine nasal cartilage proteoglycans.

作者信息

Roughley P J, Mason R M

出版信息

Biochem J. 1976 Aug 1;157(2):357-67. doi: 10.1042/bj1570357.

DOI:10.1042/bj1570357

PMID:134699

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1163861/

Abstract

Proteoglycans were extracted from bovine nasal cartilage with 2.0M-CaC2 or with 0.15M-KCl followed by 2.0M-CaC2.. Proteoglycan fractions were prepared from the extracts by density-gradient centrifugation in CsCl under 'associative' and 'dissociative' conditions. 2. The heterogeneity of the proteoglycan fractions was investigated by large-pore-gel electrophoresis. It was concluded that extracts made with 2.0M-CaCl2 or sequential 2.0M-CaCl2 contain two major species of proteoglycan 'subunit' of different hydrodynamic size, together with proteoglycan aggregates. Both 'subunits' have mobilities that are greater than those of proteoglycans obtained from pig articular cartilage McDevitt & Muir (1971) Anal. Biochem. 44, 612-622] and are therefore probably smaller in size than the latter. 3. Proteoglycan fractions isolated from cartilage extracted lith 0.15M-KCl separated into two main components on large-pore-gel electrophoresis with mobilities greater than those of proteoglycans extracted with 2.0M-CaCl2. Proteoglycans extracted at low ionic strength from bovine nasal cartilage are of similar hydrodynamic size to those extracted from pig articular cartilage under the same conditions [McDevitt & Muir (1971) Anal. Biochem. 44, 612-622]. 4. The role of endogenous proteolytic enzymes in producing proteoglycan heterogeneity, particularly in low-ionic-strength cartilage extracts is discussed. 5. Hyaluronic acid and 'link proteins' were present in the proteoglycan fraction separated from KCl extracts as well as in the fraction separated from CaCl2 extracts. Hyaluronic acid can only be identified in proteoglycan fractions by large-pore-gel electrophoresis after proteolysis and further purification of the fraction. 6. Collagen was extracted by both salt solutions and was tentatively identified as type II. Small amounts of collagen appear to be associated with the proteoglycan-aggregate fraction from the high-ionic-strength extract but not with the corresponding fraction from the KCl extract.

摘要

用2.0M - CaCl₂或先用0.15M - KCl再用2.0M - CaCl₂从牛鼻软骨中提取蛋白聚糖。通过在CsCl中“缔合”和“解离”条件下的密度梯度离心从提取物中制备蛋白聚糖级分。2. 通过大孔凝胶电泳研究蛋白聚糖级分的异质性。得出的结论是，用2.0M - CaCl₂制备的提取物或依次用2.0M - CaCl₂制备的提取物含有两种不同流体力学大小的主要蛋白聚糖“亚基”，以及蛋白聚糖聚集体。两种“亚基”的迁移率均大于从猪关节软骨中获得的蛋白聚糖的迁移率[麦克德维特和缪尔（1971年）《分析生物化学》44卷，612 - 622页]，因此其大小可能比后者小。3. 从用0.15M - KCl提取的软骨中分离得到的蛋白聚糖级分在大孔凝胶电泳上分离为两个主要成分，其迁移率大于用2.0M - CaCl₂提取的蛋白聚糖的迁移率。在低离子强度下从牛鼻软骨中提取的蛋白聚糖与在相同条件下从猪关节软骨中提取的蛋白聚糖具有相似的流体力学大小[麦克德维特和缪尔（1971年）《分析生物化学》44卷，612 - 622页]。4. 讨论了内源性蛋白水解酶在产生蛋白聚糖异质性中的作用，特别是在低离子强度软骨提取物中的作用。5. 透明质酸和“连接蛋白”存在于从KCl提取物中分离的蛋白聚糖级分以及从CaCl₂提取物中分离的级分中。透明质酸只有在蛋白水解并进一步纯化级分后才能通过大孔凝胶电泳在蛋白聚糖级分中鉴定出来。6. 两种盐溶液都能提取胶原蛋白，并初步鉴定为II型。少量胶原蛋白似乎与高离子强度提取物中的蛋白聚糖聚集体级分相关，但与KCl提取物中的相应级分无关。