title: A fully synthetic and biochemically validated phosphatidyl inositol-3-phosphate hapten via asymmetric synthesis and native chemical ligation.
abstract: We report the synthesis and biochemical validation of a phosphatidyl inositol-3 phosphate (PI3P) immunogen. The inositol stereochemistry was secured through peptide-catalyzed asymmetric phosphorylation catalysis, and the subsequent incorporation of a cysteine residue was achieved by native chemical ligation (NCL). Conjugation of the PI3P hapten to maleimide-activated keyhole limpet hemocyanin (KLH) provided a PI3P immunogen, which was successfully used to generate selective PI3P antibodies. The incorporation of a sulfhydryl nucleophile into a phosphoinositide hapten demonstrates a general strategy to reliably access phosphoinositide immunogens.
authors.affiliation: Department of Chemistry, Yale University , P.O. Box 208107, United States.
authors.forename: Brent D
authors.initials: BD
authors.lastname: Chandler
authors.forename: Anne L
authors.initials: AL
authors.lastname: Burkhardt
authors.forename: Klaudia
authors.initials: K
authors.lastname: Foley
authors.forename: Courtney
authors.initials: C
authors.lastname: Cullis
authors.forename: Denise
authors.initials: D
authors.lastname: Driscoll
authors.forename: Natalie
authors.initials: N
authors.lastname: Roy D'Amore
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Miller
mesh_terms: D003545:Cysteine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006241:Haptens / Q000737:chemistry*; D007295:Inositol Phosphates / Q000138:chemical synthesis / Q000737:chemistry*; D010716:Phosphatidylinositols / Q000737:chemistry*
chemical_list: D006241:Haptens; D007295:Inositol Phosphates; D010716:Phosphatidylinositols; D003545:Cysteine