Low Buoyant Density Proteoglycans from Saline and Dissociative Extracts of Embryonic Chicken Retinas
Low Buoyant Density Proteoglycans from Saline and Dissociative Extracts of Embryonic Chicken Retinas
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来自鸡胚视网膜盐水和解离提取物的低浮力密度蛋白多糖
DOI：
10.1111/j.1471-4159.1984.tb02752.x
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发表时间：
1984
影响因子：
4.7
通讯作者：
Anne Birkholz‐Lambrecht
中科院分区：
文献类型：
--
作者：
J. Morris;Y. Ting;Anne Birkholz‐Lambrecht
摘要：用[~3H]氨基葡萄糖或[~3H]亮氨酸和[~(35)S]硫酸盐标记培养的视网膜，然后用生理平衡盐水和4-甲基胍盐酸盐依次提取。将它们透析成缔合条件(0.5M氯化钠)，并在琼脂糖柱上进行层析。在这些条件下，一些蛋白多糖结合在大量的复合体中，当在CsCl2密度梯度中在解离条件下(4M盐酸胍)离心时，这些复合体显示出低的浮力密度。这些复合体中的大部分标记都是在蛋白多糖以外的分子中。然而，大多数蛋白多糖被包括在琼脂糖柱上，在那里它们似乎具有低浮力密度。他们抵制了通过直接氯化铯密度梯度离心法或通过二乙氨基乙基-Sephacel分离生理盐水或8M尿素提取物从主要蛋白多糖中分离潜在的低浮力密度污染物的尝试。二乙氨基乙基-Sephacel组分在碱裂解前后都经过氯化铯密度梯度离心法或Sephacryl S-300柱层析，以确认典型的O-连接糖胺聚糖的存在。中等和平衡的盐萃取物富含硫酸软骨素和其他硫酸盐大分子，可能是高度硫酸盐化的低聚糖，它们抵抗软骨素酶ABC的消化，但在电泳性上不如硫酸乙酰肝素。残渣的盐酸胍或尿素提取物是富含硫酸乙酰肝素的高密度和低密度蛋白多糖的混合物。
Abstract: Retinas were labeled in culture with [3H]glucosamine or [3H]leucine and [35S]sulfate and extracted sequentially with physiologically balanced saline and 4 M guanidine HCl. They were dialyzed into associative conditions (0.5 M NaCl) and chromatographed on agarose columns. Under these conditions, some of the proteoglycans were associated in massive complexes that showed low buoyant densities when centrifuged in CsCl density gradients under dissociative conditions (4 M guanidine HCl). Much of the label in these complexes was in molecules other than proteoglycans. Most of the proteoglycans, however, were included on the agarose columns, where they appeared to be constitutionally of low buoyant density. They resisted attempts to separate potential low buoyant density contaminants from the major proteoglycans by direct CsCl density gradient centrifugation or by the fractionation of saline or 8 M urea extracts on diethylaminoethyl‐Sephacel. The diethylaminoethyl‐Sephacel fractions were either subjected to CsCl density gradient centrifugation or were chromatographed on Sephacryl S‐300, in both cases before and after alkaline cleavage, to confirm the presence of typical O‐linked glycosaminoglycans. The medium and balanced salt extracts were enriched in chondroitin sulfate and other sul‐fated macromolecules, possibly highly sulfated oligosaccharides, that resisted digestion by chondroitinase ABC but were electrophoretically less mobile than heparan sulfate. Guanidine HCl or urea extracts of the residues were mixtures of high and low density proteoglycans that were enriched in heparan sulfate.
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DOI：
--
发表时间：
1981
期刊：
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影响因子：
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作者：
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DOI：
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发表时间：
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期刊：
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影响因子：
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作者：
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DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
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作者：
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Margolis,RK