title: Characterization of the conformations of antigenic peptides of protein lactate dehydrogenase (LDH-C4) by electrospray ionization mass spectrometry.
abstract: The conformations of several rationally designed antigenic peptides that mimic, to varying degrees, an antibody-binding region of protein lactate dehydrogenase isozyme (LDH-C4) are investigated by deuterium/hydrogen exchange and electrospray ionization mass spectrometry (ESI-MS). The approach involves monitoring the reverse-exchange of deuterium, incorporated at the labile sites in the peptides, with hydrogen as a function of time by ESI-MS. Idealized forms of a segment of the native antigen are shown to be more conformationally restricted than the native peptide based on level of deuterium that remains incorporated at the labile sites over time. From the number of amide groups of the peptide backbone that retain deuterium, estimates of the helical content of each peptide have been measured that are in close agreement with those determined by Fourier transform infrared (FTIR) spectroscopy in separate experiments. A single amino acid substitution in the idealized helical construct results in a conformational change easily detected by the deuterium exchange ESI-MS method. The approach is shown to be a viable method for characterizing the conformations of protein antigens at the local level and for screening the conformations of antigenic peptides designed to elicit optimal immune responses.
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461-1602, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Downard
mesh_terms: D000595:Amino Acid Sequence; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000276:immunology; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry* / Q000276:immunology; D011487:Protein Conformation; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D007527:Isoenzymes; D010455:Peptides; D007770:L-Lactate Dehydrogenase