Biochemical and functional characterization of proteoglycans isolated from basophils of patients with chronic myelogenous leukemia.
Human basophils were obtained from three donors with myelogenous leukemia. Proteoglycans were labeled by using [35S]sulfate as precursor and were extracted in 1 M NaCl with protease inhibitors to preserve their native structure. [35S]proteoglycans filtered on Sepharose 4B with an average m.w. similar to that of a rat heparin proteoglycan that has an estimated m.w. of 750,000. The [35S]glycosaminoglycan side chains filtered with an average m.w. slightly smaller than a 60,000-m.w. glycosaminoglycan marker. The [35S]glycosaminoglycans were resistant to heparinase and susceptible to degradation by chondroitin AC lyase and chondroitin ABC lyase. The intact [35S]glycosaminoglycans chromatographed on DEAE Sepharose as a single peak eluting just before an internal heparin marker. These findings indicate that the [35S]glycosaminoglycans were made up only of chondroitin sulfates. No heparin was identified. The chondroitin sulfate disaccharides that resulted from the action of chondroitin ABC lyase on the basophil glycosaminoglycans consisted of 92% delta Di-4S, 6% delta Di-6S, and 2% disulfated disaccharides. The [35S]chondroitin sulfate proteoglycans were susceptible to cleavage with proteases and could be shown to be released intact from basophils during degranulation initiated by the calcium ionophore A23187. The basophil proteoglycans and glycosaminoglycans were capable of binding histamine in water, but not in phosphate-buffered saline, and had no anticoagulant activity.
D001491:Basophils / Q000032:analysis* / Q000276:immunology; D001777:Blood Coagulation / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006636:Histamine Release; D006801:Humans; D007951:Leukemia, Myeloid / Q000276:immunology*; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011509:Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology
D011508:Chondroitin Sulfate Proteoglycans; D006025:Glycosaminoglycans; D011509:Proteoglycans; D000001:Calcimycin; D011133:Polysaccharide-Lyases