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ISOTOPE-CODED IONIZATION-ENHANCING REAGENTS (ICIER) FOR HIGH-THROUGHPUT PROTEIN IDENTIFICATION AND QUANTITATION USING MATRIX-ASSISTED LASER DESORPTION IONIZATION MASS SPECTROMETRY
ISOTOPE-CODED IONIZATION-ENHANCING REAGENTS (ICIER) FOR HIGH-THROUGHPUT PROTEIN IDENTIFICATION AND QUANTITATION USING MATRIX-ASSISTED LASER DESORPTION IONIZATION MASS SPECTROMETRY
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机译:基质辅助激光解吸电离质谱法同位素分析电离增强剂(ICIER),用于高通量蛋白质鉴定和定量
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摘要
Arginine-containing cysteine-modifying compounds useful for MALDI-MS analysis of proteins are provided. These compounds termed isotope-coded ionization enhancement reagents (ICIER) can provide ionization enhancement in MALDI-MS, relative quantitation, and additional database searching constraints at the same time without any extra sample manipulation. More specifically, ICIER increase the ionization efficiency of cysteine-containing peptides by attachment of a guanidino functional group. ICIER also increase the overall hydrophilicity of these peptides due the hydrophilic nature of ICIER and thus increase the percentage of recovery of these peptides during sample handling and processing such as in-gel digestion or liquid chromatography. Finally, a combination of both light and heavy ICIER provides an accurate way to obtain relative quantitation of proteins by MALDI-MIS and additional database searching constraints (number of cysteine residues in every single peptide peak) to increase the confidence of protein identification by peptide mass mapping.
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