Isolation of the major chondroitin sulfate/dermatan sulfate and heparan sulfate proteoglycans from embryonic chicken retina.
Isolation of the major chondroitin sulfate/dermatan sulfate and heparan sulfate proteoglycans from embryonic chicken retina.
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从鸡胚视网膜中分离主要的硫酸软骨素/硫酸皮肤素和硫酸乙酰肝素蛋白聚糖。
DOI：
10.1016/0003-9861(84)90261-3
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发表时间：
1984
影响因子：
3.9
通讯作者：
John E. Morris
中科院分区：
文献类型：
--
作者：
John E. Morris
本文介绍了一种从14天鸡胚视网膜中制备三种主要蛋白聚糖的技术，这些蛋白聚糖是用[35 S]硫酸盐和[3 H]葡糖胺或[3 H]丝氨酸培养过夜后得到的。组织在盐水中的均质化允许从大多数硫酸乙酰肝素蛋白聚糖中分别提取异质可溶性蛋白聚糖。后者用溶于8 ml尿素的0.5%Triton X-100从140，000 g沉淀中提取。将培养基加生理盐水和尿素洗涤剂浸提液与低分子量污染物分离，并在Sephacryl S-300（含3 murea和0.5% Triton X-100的生理盐水）上分馏成两个放射性峰。在DEAE-Sephacel上直接从这些组分中分离蛋白多糖，并进行超滤浓缩，然后在4 M盐酸胍中进行氯化铯密度梯度离心进一步纯化。检查了涉及氯化十六烷基吡啶鎓沉淀的进一步步骤，但其基本上不导致进一步纯化。分级分离：（a）从培养基中分离出一种大的硫酸软骨素/硫酸皮肤素蛋白聚糖，该蛋白聚糖被S-300排除在外，具有低浮力密度;（B）从尿素-洗涤剂提取物中分离出一种大的硫酸乙酰肝素蛋白聚糖，该蛋白聚糖也被S-300排除在外，具有低浮力密度;以及（c）两种较小的可能相关的硫酸乙酰肝素蛋白聚糖。一种是在培养基中发现的，并显示出低至中等的浮力密度;另一种是从尿素洗涤剂提取物中分离出来的，并显示出明显较高的浮力密度，与较低的蛋白质含量相关。生理盐水浸提液含有两种较大的蛋白聚糖，仅含有少量的较小分子。
A technique is presented for the preparation of three major proteoglycans from 14-day embryonic chicken retinas following their culture overnight with [35S]sulfate and either [3H]glucosamine or [3H]serine. Homogenization of the tissue in saline permitted extraction of heterogeneous soluble proteoglycans separately from most of the heparan sulfate proteoglycans. The latter were extracted from the 140,000gpellet with 0.5% Triton X-100 in 8murea. The medium plus the saline and urea-detergent extracts were separated from low-molecular-weight contaminants, and fractionated into two peaks of radioactivity on Sephacryl S-300 in saline with 3murea and 0.5% Triton X-100. The proteoglycans were isolated directly from these fractions on DEAE-Sephacel, and subjected to ultrafiltration concentration and then further purification on cesium chloride density gradient centrifugation in 4mguanidine hydrochloride. A further step involving cetylpyridinium chloride precipitation was examined, but it resulted in essentially no further purification. The fractionations separated (a) a large chondroitin sulfate/dermatan sulfate proteoglycan from the culture medium that was excluded from S-300 and of low buoyant density; (b) a large heparan sulfate proteoglycan from the urea-detergent extract that was also excluded from S-300 and of low buoyant density; and (c) two smaller and possibly related heparan sulfate proteoglycans. One was found in the medium and showed low to intermediate buoyant density; the other was isolated from the urea-detergent extract and showed a significantly higher buoyant density, associated with a lower protein content. The saline extract contained both of the two larger proteoglycans and only minor amounts of the smaller molecules.
硫酸软骨素蛋白多糖和脑可溶性糖蛋白的分级和特性。
DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kiang,WL;Margolis,RU;Margolis,RK
通讯作者：
Margolis,RK