Co-sedimentation of chondroitin sulfate A glycosaminoglycans and proteoglycans with the cytolytic secretory granules of rat large granular lymphocyte (LGL) tumor cells, and identification of a mRNA in normal and transformed LGL that encodes proteoglycans.
Rat large granular lymphocyte (LGL) tumor cell lines were analyzed for the presence of proteoglycans and glycosaminoglycans in their cytolytic secretory granules. When isolated rat LGL tumor cells were incubated in vitro for 1 to 3 hr with [35S]sulfate, and the 35S-labeled macromolecules were purified by density-gradient centrifugation, they filtered on Sepharose CL-4B columns predominantly as approximately 500,000 m.w. macromolecules. After 19 hr of incubation with [35S]sulfate, however, an 85,000 m.w. species predominated. Pulse-chase experiments revealed that the larger macromolecules were proteoglycans that with time were processed to glycosaminoglycan-sized macromolecules. As assessed by their susceptibility to chemical and enzymatic degradation and by high pressure liquid chromatography of the chondroitinase ABC-generated unsaturated disaccharides, the cell-associated rat LGL tumor cell proteoglycans bore almost exclusively chondroitin sulfate A glycosaminoglycans. Northern blot analysis using a gene-specific probe revealed that both normal peripheral blood and transformed rat LGL expressed the same approximately 1.3-kb mRNA that encodes the peptide core of the proteoglycans in the secretory granules of rat and mouse mast cells. In vivo radiolabeling of rat LGL tumor cells and isolation of their intact granules after nitrogen cavitation and density sedimentation established that glycosaminoglycans compartmentalized with cytolytic activity. Thus these negatively charged macromolecules may play a role in the regulation of the packaging and delivery of the cytolysins and basically charged serine proteases that have been identified in the cytolytic secretory granules of LGL.
D053674:Aggrecans; D000818:Animals; D002460:Cell Line; D002809:Chondroitin Sulfates / Q000235:genetics / Q000302:isolation & purification; D003594:Cytoplasmic Granules / Q000032:analysis*; D004247:DNA / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000235:genetics; D006025:Glycosaminoglycans / Q000235:genetics / Q000302:isolation & purification*; D007694:Killer Cells, Natural / Q000032:analysis* / Q000648:ultrastructure; D037181:Lectins, C-Type; D007942:Leukemia, Experimental / Q000235:genetics / Q000473:pathology; D012515:Mast-Cell Sarcoma / Q000235:genetics; D008649:Mesonephroma / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics / Q000302:isolation & purification; D011509:Proteoglycans / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000032:analysis*; D012334:RNA, Neoplasm / Q000032:analysis*; D051381:Rats; D011916:Rats, Inbred F344
C508320:Acan protein, mouse; C508321:Acan protein, rat; D053674:Aggrecans; D004273:DNA, Neoplasm; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D006025:Glycosaminoglycans; D037181:Lectins, C-Type; D009363:Neoplasm Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C038142:chondroitin sulfate glycosaminoglycan; D002809:Chondroitin Sulfates; D004247:DNA