Fluorescent detection of alpha-aminoadipic and gamma-glutamic semialdehydes in oxidized proteins.
The oxidative modification of proteins is believed to play a critical role in the etiology and/or progression of several diseases. alpha-Aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS) residues represent major oxidized amino acids generated in oxidized proteins. This paper describes a novel procedure for the specific and sensitive determination of AAS and GGS after their reductive amination with sodium cyanoborohydride and p-aminobenzoic acid, a fluorescence reagent, to their corresponding derivatives, followed by a high-performance liquid chromatography (HPLC) analysis. This fluorescent labeling of protein-associated aldehyde moieties is a simple and accurate technique that may be widely used to reveal increased levels of oxidatively modified proteins with reactive oxygen species during aging and disease.
Akagawa
Mitsugu
M
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai 599-8531, Japan.
Suyama
Kyozo
K
Uchida
Koji
K
D015074:2-Aminoadipic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D010129:4-Aminobenzoic Acid / Q000737:chemistry; D000272:Adipates / Q000378:metabolism*; D001671:Biochemistry / Q000379:methods; D001894:Borohydrides / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid / Q000031:analogs & derivatives / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017382:Reactive Oxygen Species; D012680:Sensitivity and Specificity
D000272:Adipates; D001894:Borohydrides; D005456:Fluorescent Dyes; D005971:Glutamates; D017382:Reactive Oxygen Species; D015074:2-Aminoadipic Acid; D018698:Glutamic Acid; C021199:glutamic acid gamma-semialdehyde; C029900:adipic acid; C009282:sodium cyanoborohydride; D010129:4-Aminobenzoic Acid