title: Polyamine binding to proteins in oat and Petunia protoplasts.
abstract: Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants.  To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa).  In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action.  In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH.  The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS.  Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons.  Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.
authors.affiliation: Department of Biology, Yale University, New Haven, Connecticut 06511, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Mizrahi
authors.forename: P B
authors.initials: PB
authors.lastname: Applewhite
authors.forename: A W
authors.initials: AW
authors.lastname: Galston
mesh_terms: D018554:Avena / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D003959:Diamines / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008027:Light; D010940:Plant Proteins / Q000378:metabolism*; D011073:Polyamines / Q000378:metabolism*; D011485:Protein Binding; D011523:Protoplasts / Q000378:metabolism* / Q000648:ultrastructure; D019657:Solanaceae / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D013095:Spermidine / Q000378:metabolism; D013696:Temperature
chemical_list: D003959:Diamines; D010940:Plant Proteins; D011073:Polyamines; C009475:trimethylenediamine; D013095:Spermidine