Work

Quantitative Protein Analysis Using Enzymatic [18O]Water Labeling

Year: 2014

Type: article

Abstract: This unit describes the stepwise procedure for differential oxygen isotope labeling of peptides and mass spectrometric quantification of relative protein levels in comparative proteomic experiments. The [¹⁸O] labeling of peptides happens at the peptide C-terminus and is achieved via the enzymatic oxygen exchange of tryptic peptides via catalysis of immobilized trypsin. Experimental considerations in effective incorporation and stabilization of the oxygen labels are discussed. Methods for mass spectrometric quantification of peptides with differential [¹⁶O] and [¹⁸O] isotopes are presented.

Source: Current Protocols in Protein Science

DOI: 10.1002/0471140864.ps2304s76

Authors: Mary Joan Castillo, Kristy J. Reynolds, Alexander Gomes, Catherine Fenselau, Xudong Yao

Institutions: University of Connecticut, University of Maryland, College Park

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: National Institutes of Health, National Cancer Institute

Awards: Ultrathroughput Multiple Reaction Monitoring Mass Spectrometry for Large-Scale Ca, G6407180843