Oegema T R, Bradford D S, Cooper K M
J Biol Chem. 1979 Nov 10;254(21):10579-81.
Proteoglycans isolated under associative conditions in the presence of protease inhibitors from human nucleus pulposus contained 17% aggregate and 83% non-aggregating monomer (Kav = 0.5 on Sepharose CL-2B). Isolated aggregate after reduction and alkylation was resolved into two components (Kav = 0.15 and 0.43) on Sepharose CL-2B. Labeled proteoglycans isolated from parallel samples pulsed with [35S]sulfate and chased for up to 18 h were present largely as aggregated material (up to 78%). Reduction and alkylation of the labeled samples gave a labeled proteoglycan monomer with Kav = 0.15. Both the labeled and unlabeled chondroitin sulfate chains had the same distribution on Sepharose CL-6B and equivalent molecular weights (Mr = 2.0 x 10(3)). After chondroitinase ABC digestion, the unlabeled keratan sulfate-protein core was polydisperse with a Kav = 0.38 on Sepharose CL-4B while the labeled keratan sulfate-protein core had a Kav = 0.05. This indicates that the newly synthesized proteoglycan had a large core protein and suggests that the proteoglycans present in nucleus pulposus are originally synthesized as large molecular weight, aggregating proteoglycans.
在蛋白酶抑制剂存在的条件下，于缔合状态下从人髓核中分离得到的蛋白聚糖含有17%的聚集体和83%的非聚集单体（在琼脂糖CL-2B上的Kav = 0.5）。还原和烷基化后的分离聚集体在琼脂糖CL-2B上被解析为两个组分（Kav = 0.15和0.43）。从用[35S]硫酸盐脉冲并追踪长达18小时的平行样品中分离得到的标记蛋白聚糖主要以聚集物质形式存在（高达78%）。标记样品的还原和烷基化产生了Kav = 0.15的标记蛋白聚糖单体。标记和未标记的硫酸软骨素链在琼脂糖CL-6B上具有相同的分布和相当的分子量（Mr = 2.0×10(3)）。经软骨素酶ABC消化后，未标记的硫酸角质素-蛋白核心在琼脂糖CL-4B上具有多分散性，Kav = 0.38，而标记的硫酸角质素-蛋白核心的Kav = 0.05。这表明新合成的蛋白聚糖具有较大的核心蛋白，并提示髓核中存在的蛋白聚糖最初是以大分子量的聚集蛋白聚糖形式合成的。
