title: Influence of Lβ-, Dα- and Dβ-Asp isomers of the Asp-76 residue on the properties of αA-crystallin 70-88 peptide.
abstract: Proteins have been considered to consist exclusively of L-amino acids in living tissues. However, our previous studies showed that two specific aspartyl (Asp) residues in αA- and αB-crystallins from human eye lenses invert to the D-isomers to a high degree during aging. The reaction is also accompanied by isomerization into a form containing β-Asp (isoaspartate) residues. The appearance of D- and β-Asp in a protein potentially induces large changes to the higher order structure of the protein as well as to its function. However, it remains unclear whether the formation of the Asp isomer is the direct trigger of the change to the higher order structure and function. In this study, in order to clarify the effect of the inversion to D-isomers in a protein, we synthesized peptides corresponding to the 70-88 (KFVIFLDVKHFSPEDLTVK) fragment of human αA-crystallin and its corresponding diastereoisomers in which Lα-Asp was replaced with Lβ-Asp, Dα-Asp, and Dβ-Asp at position 76 and compared their biochemical properties with that of normal peptide. The peptides containing abnormal isomers (Lβ-Asp, Dα-Asp, and Dβ-Asp residues, respectively) were more hydrophilic than the normal peptide (containing Lα-Asp), lost β-sheet structure and changed to random structures. The normal peptide promoted the aggregation of insulin while the other three isomers suppressed the aggregation of insulin. This is the first evidence that a single substitution of an Asp isomer in a peptide induces a large change to the properties of the peptide.
authors.affiliation: Research Reactor Institute, Kyoto University, Kumatori-cho, Sennan-gun, Osaka, 590-0494, Japan. nfujii@rri.kyoto-u.ac.jp
authors.forename: Noriko
authors.initials: N
authors.lastname: Fujii
authors.forename: Norihiko
authors.initials: N
authors.lastname: Fujii
authors.forename: Masashi
authors.initials: M
authors.lastname: Kida
authors.forename: Tadatoshi
authors.initials: T
authors.lastname: Kinouchi
mesh_terms: D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D006801:Humans; D007536:Isomerism; D007908:Lens, Crystalline / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D038202:alpha-Crystallin A Chain / Q000737:chemistry*
chemical_list: D010446:Peptide Fragments; D038202:alpha-Crystallin A Chain; D001224:Aspartic Acid