title: Synthesis of reagents for the construction of hypusine and deoxyhypusine peptides and their application as peptidic antigens.
abstract: Two new synthetic methods which allow access to (2S)-deoxyhypusine, natural (2S,9R)-hypusine, (2S,9S)-hypusine, and deoxyhypusine- and hypusine-containing peptides are described. The methods involve both the construction of a deoxyhypusine reagent in which the alpha-nitrogen protecting group is orthogonal to the N-7 and N-12 protecting groups and an alternate synthesis of our previous hypusine reagent, a synthesis which provides for better stereochemical control at C-9. Synthetic hypusine and deoxyhypusine can be generated from these reagents. The hypusine-containing hexapeptide (Cys-Thr-Gly-Hpu-His-Gly) is conjugated to ovalbumin (OVA), keyhole limpet hemocyanin (KLH), and a bis-maleimide; KLH conjugates are also made with the deoxyhypusine- and lysine-containing hexapeptides. Monoclonal antibodies are generated to the hypusine-containing hexapeptide-OVA conjugate in mice. These are isolated and screened against the hypusine-containing hexapeptide-KLH and hypusine-containing hexapeptide-bis-maleimide conjugates, as well as against the deoxyhypusine-containing and lysine-containing hexapeptide-KLH conjugates. These antibodies may be useful in localizing intracellular hypusine-containing peptides as well as peptides containing hypusine analogues.
authors.affiliation: Department of Medicinal Chemistry, University of Florida, Gainesville, Florida 32610-0485, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: Bergeron
authors.forename: W R
authors.initials: WR
authors.lastname: Weimar
authors.forename: R
authors.initials: R
authors.lastname: Müller
authors.forename: C O
authors.initials: CO
authors.lastname: Zimmerman
authors.forename: B H
authors.initials: BH
authors.lastname: McCosar
authors.forename: H
authors.initials: H
authors.lastname: Yao
authors.forename: R E
authors.initials: RE
authors.lastname: Smith
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D016466:CHO Cells; D006224:Cricetinae; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology; D008301:Maleimides / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009842:Oligopeptides / Q000737:chemistry* / Q000276:immunology; D010047:Ovalbumin / Q000276:immunology; D010448:Peptide Initiation Factors / Q000737:chemistry / Q000276:immunology; D016601:RNA-Binding Proteins; D013237:Stereoisomerism; D000097574:Eukaryotic Translation Initiation Factor 5A
chemical_list: D000911:Antibodies, Monoclonal; D006241:Haptens; D006433:Hemocyanins; D008239:Lysine; D008301:Maleimides; D009842:Oligopeptides; D010047:Ovalbumin; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A; C100028:hypusine; C034924:deoxyhypusine; C032808:keyhole-limpet hemocyanin