Heparin-sepharose as a tool in the subcellular fractionation of a polyamine-rich organ (rat ventral prostate) |
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Authors: | Bruna Tadolini Luciana Cabrini |
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Institution: | (1) Istituto di Chimica Biologica, Università di Bologna, Via Irneriio, 48, 40126 Bologna, Italy |
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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. |
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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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