Edwards I J, Wagner W D
Department of Comparative Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103.
J Biol Chem. 1988 Jul 15;263(20):9612-20.
Proteoglycan (PG) metabolism by aortic smooth muscle cell cultures derived from atherosclerosis-susceptible White Carneau (WC) and -resistant Show Racer (SR) pigeons was compared using [35S]sodium sulfate and [3H]serine or [3H]glucosamine as labeling precursors. Chondroitin sulfate (CS) PG and dermatan sulfate (DS) PG were the major PG secreted into the medium by both cell types. Total PG production, whether measured by incorporation of radiolabel into either core protein or glycosaminoglycan chains, was consistently lower in WC compared to SR cultures at several time points. This difference was due in part to lower (30-37%) PG synthesis in WC cells, but degradation of newly synthesized PG was an important contributor. A pulse-chase study indicated that of the total radiolabeled PG present at time O, only 47% was present at 24 h in WC cultures compared to 88% in SR cultures. The large CS-PG appeared to be the primary target for degradation in WC cells, and this selective processing resulted in a higher DS-PG:CS-PG ratio in these cultures. Structural studies indicated similar core protein and glycosaminoglycan chain sizes within a PG type for both cell types. PG monomer composition differed, however, by a higher sulfation of WC CS-PG compared to SR CS-PG and by a disaccharide sulfation position favoring 6-sulfation in WC PG and 4-sulfation in SR PG.
利用[35S]硫酸钠和[3H]丝氨酸或[3H]葡糖胺作为标记前体,比较了源自易患动脉粥样硬化的白卡诺(WC)鸽和抗动脉粥样硬化的赛鸽(SR)鸽的主动脉平滑肌细胞培养物中蛋白聚糖(PG)的代谢情况。硫酸软骨素(CS)PG和硫酸皮肤素(DS)PG是这两种细胞类型分泌到培养基中的主要PG。在几个时间点,无论是通过将放射性标记掺入核心蛋白还是糖胺聚糖链来测量,WC培养物中的总PG产量始终低于SR培养物。这种差异部分是由于WC细胞中PG合成较低(30-37%),但新合成PG的降解也是一个重要因素。脉冲追踪研究表明,在时间O时存在的总放射性标记PG中,WC培养物在24小时时仅存在47%,而SR培养物中为88%。大的CS-PG似乎是WC细胞中降解的主要靶点,这种选择性加工导致这些培养物中DS-PG:CS-PG的比例更高。结构研究表明,两种细胞类型在一种PG类型内的核心蛋白和糖胺聚糖链大小相似。然而,PG单体组成不同,WC CS-PG的硫酸化程度高于SR CS-PG,并且二糖硫酸化位置有利于WC PG中的6-硫酸化和SR PG中的4-硫酸化。