Methods for assaying nonenzymatic glycosylation.

Open University, Oxford Research Unit, England.
Analytical Biochemistry (Impact Factor: 2.58). 01/1989; 175(2):347-60. DOI: 10.1016/0003-2697(88)90558-1
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    ABSTRACT: Glycation is a common modification of proteins both in vitro and in vivo. To aid identification and comparative quantitation, a method of stable isotope labeling followed by LC-MS analysis was proposed. The samples were reduced using sodium borohydride or sodium borodeuteride. Reduction of the Schiff base between the amine group and the reducing sugars resulted in a molecular weight increase of 2Da using sodium borohydride or a molecular weight increase of 3Da using sodium borodeuteride. The molecular weight difference of 1Da between peptides containing glycated lysine residue reduced using sodium borohydride or sodium borodeuteride was used to identify glycated peptides and to calculate the glycation difference between samples. The method was used to investigate glycation of a recombinant human IgG1 antibody under native and denaturing conditions. The result demonstrated a good correlation between glycation propensity of lysine residues and their solvent exposure levels.
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 03/2014; 958C:90-95. · 2.78 Impact Factor
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    IJCBS. 01/2014;
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    ABSTRACT: The use of tandem mass spectrometry to identify and characterize sites of protein adenosine diphosphate (ADP) ribosylation will be reviewed. Specifically, we will focus on data acquisition schemes and fragmentation techniques that provide peptide sequence and modification site information. Also discussed are uses of synthetic standards to aid characterization, and an enzymatic method that converts ADP-ribosylated peptides into ribosyl mono phosphorylated peptides making identification amenable to traditional phosphopeptide characterization methods. Finally the potential uses of these techniques to characterize poly ADP-ribosylation sites, and inherent challenges, are addressed.
    International Journal of Mass Spectrometry 02/2012; 312:114-121. · 2.14 Impact Factor