A novel class of chemically modified iodo-containing resins: design, synthesis and application to mass spectrometry-based proteome analysis.
A novel class of chemically modified iodo-containing resins with isotope-labeled tagging for mass spectrometry-based proteome analysis is described. This iodo-containing resin contains a thiol-reactive group that is used to capture the cysteine (Cys)-containing peptides from peptide mixtures, one 'tag' amino acid, and an aminomethyl polystyrene resin with Rink Amide Linker. The 'tag' amino acid is synthesized in both heavy and light isotope-coded forms and therefore permits the direct relative quantification of peptides/proteins through mass spectrometric analysis. In the iodo-containing resin strategy, the Cys-containing peptides of two samples covalently captured by either light or heavy iodo-containing resin were mixed and washed extensively under stringent conditions. Then the Cys-containing peptides were retrieved by acid-catalyzed elution. Finally, the eluted peptides were directly analyzed by micro liquid chromatography/mass spectrometry for identification and relative quantification. The iodo-containing resins were synthesized by a simple but effective method. Their abilities to identify and quantify the Cys-containing part in two samples were proved by the analysis of mixtures of amino acids, peptides and proteins.
Zhang
Li
L
Shanghai Mass Spectrometry Center, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.
Guo
Yin-Long
YL
Liu
Han-Qing
HQ
D000595:Amino Acid Sequence; D003545:Cysteine / Q000032:analysis; D015195:Drug Design; D007454:Iodides / Q000737:chemistry*; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011137:Polystyrenes / Q000138:chemical synthesis / Q000737:chemistry; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D012117:Resins, Synthetic / Q000138:chemical synthesis* / Q000737:chemistry*
D007454:Iodides; D010455:Peptides; D011137:Polystyrenes; D020543:Proteome; D012117:Resins, Synthetic; D009113:Muramidase; D003545:Cysteine