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Click Chemistry Facilitates Formation of Reporter Ions and Simplified Synthesis of Amine-Reactive Multiplexed Isobaric Tags for Protein Quantification

Sohn, Chang Ho and Lee, J. Eugene and Sweredoski, Michael J. and Graham, Robert L. J. and Smith, Geoffrey T. and Hess, Sonja and Czerwieniec, Gregg and Loo, Joseph A. and Deshaies, Raymond J. and Beauchamp, J. L. (2012) Click Chemistry Facilitates Formation of Reporter Ions and Simplified Synthesis of Amine-Reactive Multiplexed Isobaric Tags for Protein Quantification. Journal of the American Chemical Society, 134 (5). pp. 2672-2680. ISSN 0002-7863. PMCID PMC3668889. https://resolver.caltech.edu/CaltechAUTHORS:20120202-074303275

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Abstract

We report the development of novel reagents for cell-level protein quantification, referred to as Caltech isobaric tags (CITs), which offer several advantages in comparison with other isobaric tags (e.g., iTRAQ and TMT). Click chemistry, copper(I)-catalyzed azide–alkyne cycloaddition (CuAAC), is applied to generate a gas-phase cleavable linker suitable for the formation of reporter ions. Upon collisional activation, the 1,2,3-triazole ring constructed by CuAAC participates in a nucleophilic displacement reaction forming a six-membered ring and releasing a stable cationic reporter ion. To investigate its utility in peptide mass spectrometry, the energetics of the observed fragmentation pathway are examined by density functional theory. When this functional group is covalently attached to a target peptide, it is found that the nucleophilic displacement occurs in competition with formation of b- and y-type backbone fragment ions regardless of the amino acid side chains present in the parent bioconjugate, confirming that calculated reaction energetics of reporter ion formation are similar to those of backbone fragmentations. Based on these results, we apply this selective fragmentation pathway for the development of CIT reagents. For demonstration purposes, duplex CIT reagent is prepared using a single isotope-coded precursor, allyl-d_5-bromide, with reporter ions appearing at m/z 164 and 169. Isotope-coded allyl azides for the construction of the reporter ion group can be prepared from halogenated alkyl groups which are also employed for the mass balance group via N-alkylation, reducing the cost and effort for synthesis of isobaric pairs. Owing to their modular designs, an unlimited number of isobaric combinations of CIT reagents are, in principle, possible. The reporter ion mass can be easily tuned to avoid overlapping with common peptide MS/MS fragments as well as the low mass cutoff problems inherent in ion trap mass spectrometers. The applicability of the CIT reagent is tested with several model systems involving protein mixtures and cellular systems.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja2099003DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3668889/PubMed CentralArticle
ORCID:
AuthorORCID
Sweredoski, Michael J.0000-0003-0878-3831
Hess, Sonja0000-0002-5904-9816
Deshaies, Raymond J.0000-0002-3671-9354
Beauchamp, J. L.0000-0001-8839-4822
Additional Information:© 2012 American Chemical Society. Received: October 20, 2011. Just Accepted Manuscript January 05, 2012. Publication Date (Web): January 5, 2012. This work was supported by the National Science Foundation (NSF) through grant CHE-0416381 (J.L.B.), the Beckman Institute at California Institute of Technology (J.L.B., M.J.S., S.H., and R.J.L.G.), and the National Institutes of Health (NIH) through grant RR 20004 (J.A.L.), and the Betty and Gordon Moore Foundation (S.H. and R.J.L.G). Computational resources for DFT results were kindly provided by the Materials and Process Simulation Center at California Institute of Technology. C.H.S. acknowledges a fellowship from the Kwanjeong Educational Foundation. J.E.L. was supported by the Ruth L. Kirschstein NRSA fellowship from the NIH (CA138126).
Funders:
Funding AgencyGrant Number
NSFCHE-0416381
Caltech Beckman InstituteUNSPECIFIED
NIHRR 20004
Gordon and Betty Moore FoundationUNSPECIFIED
Kwanjeong Educational FoundationUNSPECIFIED
NIH Predoctoral FellowshipCA138126
Issue or Number:5
PubMed Central ID:PMC3668889
Record Number:CaltechAUTHORS:20120202-074303275
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20120202-074303275
Official Citation:Click Chemistry Facilitates Formation of Reporter Ions and Simplified Synthesis of Amine-Reactive Multiplexed Isobaric Tags for Protein Quantification Chang Ho Sohn, J. Eugene Lee, Michael J. Sweredoski, Robert L.J. Graham, Geoffrey T. Smith, Sonja Hess, Gregg Czerwieniec, Joseph A. Loo, Raymond J. Deshaies, and J. L. Beauchamp Journal of the American Chemical Society 2012 134 (5), 2672-2680
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29080
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:13 Mar 2012 22:24
Last Modified:03 Oct 2019 03:38

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