Design and synthesis of a solid-phase fluorescent mass tag

✍ Scribed by Yang Shi; Rong Xiang; Csaba Horváth; James A. Wilkins

• Publisher: John Wiley and Sons
• Year: 2005
• Tongue: English
• Weight: 393 KB
• Volume: 28
• Category: Article
• ISSN: 1615-9306
• DOI: 10.1002/jssc.200500115

✦ Synopsis

Abstract

In this study, we demonstrate the design of a new solid‐phase fluorescent mass tag (FMT) that contains the following features: (1) the FMT is synthesized using Fmoc chemistry which is simple, rapid, and cost‐effective; (2) lysine is used as a uniformly labeled amino acid (using stable isotopes) to allow 8 Da difference between “heavy” and “light” tags; (3) a fluorescent molecule is coupled to the isotope tag that allows a tagged peptide to be detected by online fluorescence; and (4) an iodoacetyl reactive group provides cysteine reactivity. Using MALDI‐TOF MS and HPLC, we show that the FMT reagent can be used to label standard cysteine‐containing peptides as well as cysteine‐containing peptides from a BSA tryptic digest.