Gardiner E E, Mok S S, Sriratana A, Robinson H C, Veitch B J, Lowther D A, Handley C J
Department of Biochemistry, Monash University, Clayton, Australia.
Eur J Biochem. 1994 Apr 15;221(2):871-9. doi: 10.1111/j.1432-1033.1994.tb18802.x.
Rabbit peritoneal polymorphonuclear neutrophils (PMN), incubated in medium containing [35S]sulphate, incorporated 35S into proteoglycan and protein fractions. Approximately 46% of the 35S-labelled macromolecules associated with the PMN cells after 1 h of incubation were recovered in a cytoplasmic granule extract, the majority being present in azurophil granules. Analysis of the azurophil granule fraction showed that approximately 90% of the 35S-labelled macromolecules were proteoglycans. When challenged with heat-aggregated rabbit gamma-globulin in the presence of cytochalasin B and cGMP, PMN were induced to release granular enzymes but did not release 35S-labelled proteoglycans into the incubation medium. When incubated with articular cartilage slices, PMN released their granule 35S-labelled proteoglycan into the medium and into the cartilage matrix. Granule enzymes and 35S-labelled granule proteoglycan were extracted from the cartilage tissue after incubation and 35S-labelled macromolecules were detected in the cartilage tissue by autoradiography.
将兔腹膜多形核中性粒细胞(PMN)置于含有[35S]硫酸盐的培养基中孵育,35S会掺入蛋白聚糖和蛋白质组分中。孵育1小时后,与PMN细胞相关的35S标记大分子中约46%在细胞质颗粒提取物中回收,大部分存在于嗜天青颗粒中。对嗜天青颗粒部分的分析表明,35S标记大分子中约90%是蛋白聚糖。当在细胞松弛素B和环鸟苷酸存在的情况下用热聚集兔γ球蛋白刺激时,PMN被诱导释放颗粒酶,但未将35S标记的蛋白聚糖释放到孵育培养基中。当与关节软骨切片一起孵育时,PMN将其颗粒35S标记的蛋白聚糖释放到培养基和软骨基质中。孵育后从软骨组织中提取颗粒酶和35S标记的颗粒蛋白聚糖,并通过放射自显影在软骨组织中检测到35S标记的大分子。