Heparin-sepharose as a tool in the subcellular fractionation of a polyamine-rich organ (rat ventral prostate).
Heparin-sepharose forms complexes with putrescine, spermidine, and spermine and indirect measurements of its affinity for polyamines gives values similar to those obtained with free heparin. A direct measurement of the binding of heparin-sepharose to spermine gives an apparent dissociation constant (Kd) of 1.5 X 10(-6)M spermine. Unlike free heparin, heparin-sepharose does not cause either disruption of the nuclei or more sutble modifications able to modify their sedimentation behavior. The heparin-sepharose polyamine complex formed by the addition of heparin-sepharose to the homogenate can easily be removed and the homogenate can be processed according to normal schedules. Heparin-sepharose is able to sequester 85% of the exchangeable spermine present in the homogenate of rat ventral prostate. The distribution of the marker enzyme galactosyltransferase (Golgi apparatus) on a sucrose density gradient was followed to assess the usefulness of heparin-sepharose in minimizing the aggregation of cellular organelles brought about by polyamines.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000648:ultrastructure; D006493:Heparin; D006801:Humans; D008297:Male; D011073:Polyamines / Q000032:analysis*; D011467:Prostate / Q000648:ultrastructure*; D011700:Putrescine / Q000032:analysis; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives; D013095:Spermidine / Q000032:analysis; D013096:Spermine / Q000032:analysis
D011073:Polyamines; C037405:heparin-sepharose; D013096:Spermine; D006493:Heparin; D012685:Sepharose; D013095:Spermidine; D011700:Putrescine