title: Development of a hypersensitive periodate-cleavable amino acid that is methionine- and disulfide-compatible and its application in MHC exchange reagents for T cell characterisation.
abstract: Incorporation of cleavable linkers into peptides and proteins is of particular value in the study of biological processes. Here we describe the synthesis of a cleavable linker that is hypersensitive to oxidative cleavage as the result of the periodate reactivity of a vicinal amino alcohol moiety. Two strategies directed towards the synthesis of a building block suitable for solid-phase peptide synthesis were developed: a chemoenzymatic route, involving L-threonine aldolase, and an enantioselective chemical route; these led to α,γ-diamino-β-hydroxybutanoic acids in diastereoisomerically mixed and enantiopure forms, respectively. Incorporation of the 1,2-amino alcohol linker into the backbone of a peptide generated a conditional peptide that was rapidly cleaved at very low concentrations of sodium periodate. This cleavable peptide ligand was applied in the generation of MHC exchange reagents for the detection of antigen-specific T cells in peripheral blood cells. The extremely low concentration of periodate required to trigger MHC peptide exchange allowed the co-oxidation of methionine and disulfide residues to be avoided. Conditional MHC reagents hypersensitive to periodate can now be applied without limitations when UV irradiation is undesired or less practical.
authors.affiliation: Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the Netherlands.
authors.forename: Alessia
authors.initials: A
authors.lastname: Amore
authors.forename: Kim
authors.initials: K
authors.lastname: Wals
authors.forename: Evelyn
authors.initials: E
authors.lastname: Koekoek
authors.forename: Rieuwert
authors.initials: R
authors.lastname: Hoppes
authors.forename: Mireille
authors.initials: M
authors.lastname: Toebes
authors.forename: Ton N M
authors.initials: TN
authors.lastname: Schumacher
authors.forename: Boris
authors.initials: B
authors.lastname: Rodenko
authors.forename: Huib
authors.initials: H
authors.lastname: Ovaa
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D000605:Amino Alcohols / Q000737:chemistry; D001672:Biocompatible Materials / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry*; D055808:Drug Discovery; D015789:HLA-A2 Antigen / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008715:Methionine; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000494:pharmacology*; D013237:Stereoisomerism; D013379:Substrate Specificity; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D000605:Amino Alcohols; D001672:Biocompatible Materials; D004220:Disulfides; D015789:HLA-A2 Antigen; D007202:Indicators and Reagents; D010504:Periodic Acid; D008715:Methionine