Development and application of a method for identification of isothiocyanate-targeted molecules in colon cancer cells |
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| Authors: | Noriyuki Miyoshi Takumi Yonemochi Susumu Tomono Ryuuta Fukutomi Yoshimasa Nakamura Hiroshi Ohshima |
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| Institution: | Department of Food and Nutritional Sciences, Graduate School of Nutritional and Environmental Sciences, and Global Center of Excellence Program, University of Shizuoka, Shizuoka 422-8526, Japan. |
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| Abstract: | In this study, we have developed a novel method to identify isothiocyanate (ITC)-targeted molecules using two well-studied ITCs: benzyl ITC (BITC) and phenethyl ITC (PEITC). The principle of this method is based on identifying a pattern of differences between BITC and PEITC given that they show similar chemical and biological behaviors. For method validation, dithiothreitol-reduced bovine insulin as a model molecule was incubated with either BITC or PEITC, and digested peptides were analyzed by ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-TOF-MS) and liquid chromatography quadrupole TOF-MS (LC-Q-TOF-MS). Three peptides-NYCN, FVNQHLCGSHLVE, and ALYLVCGE-were identified as being adducted with BITC or PEITC on their cysteine residues. Each set of peptides adducted with either BITC or PEITC showed retention times (RT(BITC)
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