Ward J V, Packham M A
Biochim Biophys Acta. 1979 Mar 7;583(2):196-207. doi: 10.1016/0304-4165(79)90427-6.
Rabbit platelets were labeled in vivo with 35S for characterization of platelet sulfated glycosaminoglycan. When rabbit platelets were aggregated by ADP, sulfated proteoglycan was lost from the platelet surface although no release of granule contents occurred. The sulfated proteoglycan contained in the granules of platelets pretreated with ADP was subsequently released by treatment with thrombin. The 35S-labeled proteoglycan from both sources was isolated by gel filtration and the glycosaminoglycan portion of the proteoglycan was characterized as chondroitin 4-sulfate by examining the products of digestion with hyaluronidase, chondroitinase AC and ABC, and chondro-4- and 6-sulfatases; by identification of the hexosamine as N-acetylgalactosamine; by determination of a 1 : 1 : 1 molar ratio of N-acetylgalactosamine, uronic acid and inorganic sulfate; and by cetylpyridinium chloride cellulose chromatography. In these studies, the use of 35S-labeled proteoglycan made possible detection and quantification of much smaller amounts of material than would be possible with unlabeled material. Chondroitin 4-sulfate was the only sulfated glycosaminoglycan identified in the proteoglycan lost from the platelet surface during ADP-induced aggregation and in the proteoglycan released from the granules when the platelets were exposed to thrombin.
用35S对兔血小板进行体内标记，以表征血小板硫酸化糖胺聚糖。当兔血小板被ADP聚集时，硫酸化蛋白聚糖从血小板表面丢失，尽管颗粒内容物未释放。用ADP预处理的血小板颗粒中所含的硫酸化蛋白聚糖随后经凝血酶处理而释放。通过凝胶过滤分离来自两种来源的35S标记蛋白聚糖，并通过检查用透明质酸酶、软骨素酶AC和ABC以及软骨素4-和6-硫酸酯酶消化的产物；通过将己糖胺鉴定为N-乙酰半乳糖胺；通过测定N-乙酰半乳糖胺、糖醛酸和无机硫酸盐的1:1:1摩尔比；以及通过十六烷基吡啶氯化物纤维素色谱法，将蛋白聚糖的糖胺聚糖部分表征为硫酸软骨素4-硫酸盐。在这些研究中，使用35S标记的蛋白聚糖使得检测和定量比未标记材料可能检测到的量少得多的物质成为可能。硫酸软骨素4-硫酸盐是在ADP诱导的聚集过程中从血小板表面丢失的蛋白聚糖以及当血小板暴露于凝血酶时从颗粒中释放的蛋白聚糖中鉴定出的唯一硫酸化糖胺聚糖。
