title: Polyamine-linked sepharoses: preparation and application to mammalian spermine synthase.
abstract: Seven different polyamine-linked Sepharose derivatives were prepared for the affinity chromatography of spermidine and spermine binding macromolecules: Spermine synthase from rat and hog brain was used as a model protein with a spermidine binding site. Comparative studies of the affinities of the enzymes for the seven matrixes suggested that two negative charges, three to four methylene groups apart, should be present at the decarboxylated S-adenosylmethionine binding site and should improve the binding of the enzyme to the Sepharose derivative. Two negative charges at the spermidine binding site would be expected to do the same. Three affinity matrixes linked with 1,17-diamino-4,9,14-triazaheptadecane, 1,21-diamino-4,9,13,18-tetraazaheneicosane, and 5-spermine carboxylic acid, respectively, had an affinity for spermine synthases higher than that of spermine-Sepharose, which has been used for the purification of spermine synthase. The first of these matrixes was used and proved to be effective for the purification.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Josai University, Saitama, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Shirahata
authors.forename: C L
authors.initials: CL
authors.lastname: Zhu
authors.forename: S
authors.initials: S
authors.lastname: Akatsu
authors.forename: Y
authors.initials: Y
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Samejima
mesh_terms: D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification; D002846:Chromatography, Affinity; D011073:Polyamines; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives* / Q000302:isolation & purification*; D013097:Spermine Synthase / Q000737:chemistry / Q000302:isolation & purification*; D013552:Swine
chemical_list: D002352:Carrier Proteins; D011073:Polyamines; D012685:Sepharose; D013097:Spermine Synthase