Lohmander S, Hjerpe A
Biochim Biophys Acta. 1975 Sep 8;404(1):93-109. doi: 10.1016/0304-4165(75)90151-8.
Rib cartilage from growing guinea pigs and epiphyseal cartilage from Beagle puppies were separated into three fractions, representing non-mineralized, low mineralized, and high mineralized, tissue, by centrifuging finely ground material in acetone/bromoform density gradients. Following extraction under dissociative conditions, the proteoglycans were fractionated by density gradient ultracentrifugation under associative and dissociative conditions. With the onset of mineralization, the cartilage lost approximately half its content of proteoglycans. The proteoglycans remaining in the calcified cartilage differed in composition and in size from those of nonmineralized tissue. With the increased mineral content of the tissues the ratios of protein to polysaccharide, of chondroitin sulfate to keratan sulfate, and of 4-sulfate to 6-sulfated chondroitin sulfate increased in the proteoglycan fraction. Furthermore, gel chromatograms indicated decreased proportions of very high molecular weight proteoglycans, in mineralized tissue.
将生长中的豚鼠的肋软骨和比格幼犬的骨骺软骨在丙酮/溴仿密度梯度中对精细研磨的材料进行离心,分离成三个部分,分别代表未矿化、低矿化和高矿化组织。在解离条件下提取后,通过在缔合和解离条件下的密度梯度超速离心对蛋白聚糖进行分级分离。随着矿化的开始,软骨失去了大约一半的蛋白聚糖含量。钙化软骨中残留的蛋白聚糖在组成和大小上与未矿化组织中的不同。随着组织矿含量的增加,蛋白聚糖部分中蛋白质与多糖的比例、硫酸软骨素与硫酸角质素的比例以及4-硫酸化与6-硫酸化硫酸软骨素的比例增加。此外,凝胶色谱图表明矿化组织中极高分子量蛋白聚糖的比例降低。