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Heparin-sepharose as a tool in the subcellular fractionation of a polyamine-rich organ (rat ventral prostate)
Authors:Bruna Tadolini  Luciana Cabrini
Institution:(1) Istituto di Chimica Biologica, Università di Bologna, Via Irneriio, 48, 40126 Bologna, Italy
Abstract: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 (K d) of 1.5×10−6 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.
Keywords:Heparin-sepharose  in subcellular fractionation  sepharose-heparin  in subcellular fractionation  polyamine-rich organ  subcellular fractionation  rat ventral prostate  subcellular fractionation  fractionation  subcellular  by heparin-sepharose
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