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The in vitro nuclear aggregates of polyamines
Author(s) -
Di Luccia Aldo,
Picariello Gianluca,
Iacomino Giuseppe,
Formisano Annarita,
Paduano Luigi,
D’Agostino Luciano
Publication year - 2009
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2009.06960.x
Subject(s) - putrescine , spermine , chemistry , spermidine , in vitro , sodium , phosphate , chromatography , dna , biochemistry , nuclease , polyamine , polyphosphate , gel permeation chromatography , organic chemistry , enzyme , polymer
Natural polyamines (putrescine, spermidine, and spermine) self‐assemble in a simulated physiological environment (50 m m sodium phosphate buffer, pH 7.2), generating in vitro nuclear aggregates of polyamines ( iv NAPs). These supramolecular compounds are similar in structure and molecular mass to naturally occurring cellular nuclear aggregates of polyamines, and they share the ability of NAPs to interact with and protect the genomic DNA against nuclease degradation. Three main iv NAP compounds were separated by gel permeation chromatography. Their elution was carried out with 50 m m sodium phosphate buffer supplemented with 150 m m NaCl. Freezing and thawing of selected chromatographic fractions obtained by GPC runs in which the mobile phase was sodium phosphate buffer not supplemented with NaCl yielded three different microcrystallites, specifically corresponding to the iv NAPs, all of which were able to bind DNA. In this study, we demonstrated that in vitro self‐assembly of polyamines and phosphates is a spontaneous, reproducible and inexpensive event, and provided the indications for the production of the iv NAPs as a new tool for manipulating the genomic DNA machinery.