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来自不同解剖部位的狒狒(巴氏狒狒)软骨的蛋白聚糖群体:解离的蛋白聚糖及通过密度梯度离心获得的组分的凝胶电泳分析

Proteoglycan populations of baboon (Papio papio) cartilages from different anatomical sites: gel electrophoretic analysis of dissociated proteoglycans and of fractions obtained by density gradient centrifugation.

作者信息

Stanescu V, Maroteaux P, Sobczak E

出版信息

Biochim Biophys Acta. 1980 May 7;629(2):371-81. doi: 10.1016/0304-4165(80)90109-9.

Abstract

Proteoglycans extracted by 4 M guanidinium chloride from different cartilages and vertebral discs of young baboons (Papio papio) were purified by ion-exchange chromatography and assessed by gel electrophoresis. Proteoglycans fractionated by equilibrium density gradient centrifugation under 'dissociative' conditions were similarly purified and assessed. (1) Non-fractionated proteoglycans from articular, nasal, laryngeal, tracheal, costal, vertebral plate and ear cartilage and from anulus fibrosus gave three metachromatic bands on gel electrophoresis: two broad, closely running bands (I and II) and a faster, more discrete band (III). The migrations rates of corresponding bands of various cartilages were similar, with minor differences in staining intensity. Gel electrophoresis of material fractionated on the gradient indicated the presence of bands with the same migration and appearance as those found for the unfractionated proteoglycans and also partial separation of bands (buoyant density I greater than II greater than III). (2) Non-fractionated proteoglycans from growth cartillage yielded bands I and II only. Small amounts of material corresponding to band III were detected in the upper fractions of the gradients. (3) Proteoglycans from nucleus pulposus migrated as two diffuse bands corresponding to bands I and II. The two bands were partially separated in the gradients and an additional very slow band was formed in the middle fractions of the gradients. (4) Proteoglycans from knee menisci contained a densely stained band of corresponding mobility to band III. Faint bands corresponding to band I and II were also detected. The bands were partially separated in the gradients.

摘要

用4M氯化胍从幼年狒狒(巴氏狒狒)的不同软骨和椎间盘中提取的蛋白聚糖,通过离子交换色谱法进行纯化,并通过凝胶电泳进行评估。在“解离”条件下通过平衡密度梯度离心分级分离的蛋白聚糖也进行了类似的纯化和评估。(1)来自关节软骨、鼻软骨、喉软骨、气管软骨、肋软骨、椎板软骨、耳软骨和纤维环的未分级蛋白聚糖在凝胶电泳上呈现出三条异染带:两条宽的、紧密相邻的带(I和II)以及一条迁移速度更快、更离散的带(III)。各种软骨相应条带的迁移率相似,染色强度略有差异。对梯度分级分离的材料进行凝胶电泳表明,存在与未分级蛋白聚糖中发现的条带具有相同迁移率和外观的条带,并且条带也有部分分离(浮力密度I大于II大于III)。(2)来自生长软骨的未分级蛋白聚糖仅产生条带I和II。在梯度的上层部分检测到少量对应于条带III的物质。(3)来自髓核的蛋白聚糖迁移为对应于条带I和II的两条弥散带。这两条带在梯度中部分分离,并且在梯度的中间部分形成了一条额外的非常慢的带。(4)来自膝关节半月板的蛋白聚糖含有一条与条带III迁移率相应的深染带。还检测到了对应于条带I和II的淡带。这些条带在梯度中部分分离。

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