Schick B P, Senkowski-Richardson S
Cardeza Foundation for Hematologic Research, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107.
Biochem J. 1992 Mar 15;282 ( Pt 3)(Pt 3):651-8. doi: 10.1042/bj2820651.
Synthesis of sulphated proteoglycans was compared in human erythroleukaemia (HEL) cells grown under control conditions and under stimulation by dimethyl sulphoxide (DMSO) and phorbol 12-myristate 13-acetate (PMA). Synthesis of [35S]sulphate-labelled proteoglycans by DMSO-treated cells was decreased by about 35% relative to controls, but synthesis of proteoglycans by PMA-treated cells increased 3-4-fold. Control and DMSO-treated cells secreted 65% of the newly synthesized proteoglycans, but PMA-treated cells secreted more than 90%. Sepharose CL-6B chromatography and SDS/PAGE suggested the presence of several proteoglycans in the cells and culture medium. The PMA-treated cells synthesized a low-Mr proteoglycan (Kav. 0.3( that was not present in controls and DMSO-treated cultures. The proteoglycans of the cells and medium from control, DMSO-treated and PMA-treated cultures could be separated into three fractions by octyl-Sepharose chromatography. The proteoglycans were resistant to trypsin but were degraded by Pronase and papain to fragments similar in size to the NaOH/NaBH4-generated glycosaminoglycans. The average chain length of the glycosaminoglycans (Kav. 0.20 on Sepharose CL-6B for controls) was decreased by DMSO (Kav. 0.25) and by PMA (Kav. 0.30-0.38). Chondroitin ABC lyase digestion of the proteoglycans from the medium of the control cultures produced two core proteins at Mr 31,000 and 36,000. The DMSO medium proteoglycans had only the 31,000-Mr core protein, and the PMA culture medium proteoglycans had core proteins of Mr 27,000, 31,000 and 36,000. Changes in synthesis of proteoglycans induced by DMSO or PMA may have relevance for the maturation of haematopoietic cells.
在对照条件下以及在二甲基亚砜（DMSO）和佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）刺激下培养的人红白血病（HEL）细胞中，对硫酸化蛋白聚糖的合成进行了比较。与对照相比，经DMSO处理的细胞中[35S]硫酸盐标记的蛋白聚糖合成减少了约35%，但经PMA处理的细胞中蛋白聚糖的合成增加了3至4倍。对照细胞和经DMSO处理的细胞分泌了新合成蛋白聚糖的65%，但经PMA处理的细胞分泌量超过90%。琼脂糖CL - 6B层析和SDS/PAGE表明细胞和培养基中存在几种蛋白聚糖。经PMA处理的细胞合成了一种低分子量的蛋白聚糖（洗脱体积0.3），对照细胞和经DMSO处理的培养物中不存在这种蛋白聚糖。对照、经DMSO处理和经PMA处理的培养物的细胞和培养基中的蛋白聚糖通过辛基琼脂糖层析可分为三个组分。这些蛋白聚糖对胰蛋白酶有抗性，但可被链霉蛋白酶和木瓜蛋白酶降解为与NaOH/NaBH4产生的糖胺聚糖大小相似的片段。糖胺聚糖的平均链长（对照在琼脂糖CL - 6B上的洗脱体积0.20）经DMSO处理后降低（洗脱体积0.25），经PMA处理后也降低（洗脱体积0.30 - 0.38）。对照培养物培养基中的蛋白聚糖经软骨素ABC裂解酶消化产生了分子量分别为31,000和36,000的两种核心蛋白。DMSO培养基中的蛋白聚糖只有分子量为31,000的核心蛋白，而PMA培养基中的蛋白聚糖有分子量为27,000、31,000和36,000的核心蛋白。DMSO或PMA诱导的蛋白聚糖合成变化可能与造血细胞的成熟有关。
