Cohen M P, Wu V Y, Surma M L
Biochim Biophys Acta. 1981 Dec 18;678(3):322-8. doi: 10.1016/0304-4165(81)90110-0.
Extraction of rat glomerular basement membrane, purified by osmotic lysis and sequential detergent treatment, with 8 M urea containing protease inhibitors solubilizes protein that is devoid of hydroxyproline and hydroxylysine. This material represents 8-12% of total membrane protein, elutes mainly as two high molecular weight peaks on agarose gel filtration, and is associated with glycosaminoglycans. Isolated rat renal glomeruli incorporate [35S]sulfate into basement membrane from which this non-collagenous 35S-labeled fraction can be subsequently solubilized. The radioactivity incorporated into urea-soluble glomerular basement membrane eluted primarily with the higher molecular weight peak (Mr greater than 250 000). Cellulose acetate electrophoresis after pronase digestion of the urea-soluble fraction revealed glycosaminoglycan that was resistant to digestion with Streptomyces hyaluronidase and chondroitinase ABC, sensitive to nitrous acid treatment, and contained [35S]sulfate. The findings indicate that one of the non-collagenous components of glomerular basement membrane is a proteoglycan containing heparan sulfate.
通过渗透裂解和连续去污剂处理纯化的大鼠肾小球基底膜，用含有蛋白酶抑制剂的8M尿素溶解后，可使不含羟脯氨酸和羟赖氨酸的蛋白质溶解。该物质占总膜蛋白的8-12%，在琼脂糖凝胶过滤中主要以两个高分子量峰洗脱，并与糖胺聚糖相关。分离的大鼠肾小球将[35S]硫酸盐掺入基底膜中，随后可从中溶解出这种非胶原35S标记部分。掺入尿素可溶性肾小球基底膜的放射性主要与较高分子量峰（Mr大于250000）一起洗脱。对尿素可溶性部分进行链霉蛋白酶消化后的醋酸纤维素电泳显示，糖胺聚糖对透明质酸酶和软骨素酶ABC消化有抗性，对亚硝酸处理敏感，并含有[35S]硫酸盐。这些发现表明，肾小球基底膜的一种非胶原成分是含有硫酸乙酰肝素的蛋白聚糖。
