Comparison of MS(2)-only, MSA, and MS(2)/MS(3) Methodologies for Phosphopeptide Identification

Overview

Journal J Proteome Res

Publisher American Chemical Society

Specialty Biochemistry

Date 2008 Dec 17

PMID 19072539

Citations 22

Authors

Peter J Ulintz

Anastasia K Yocum

Bernd Bodenmiller

Ruedi Aebersold

Philip C Andrews

Alexey I Nesvizhskii

Affiliations

Soon will be listed here.

Abstract

Current mass spectrometers provide a number of alternative methodologies for producing tandem mass spectra specifically for phosphopeptide analysis. In particular, generation of MS(3) spectra in a data-dependent manner upon detection of the neutral loss of a phosphoric acid in MS(2) spectra is a popular technique for circumventing the problem of poor phosphopeptide backbone fragmentation. The newer Multistage Activation method provides another option. Both these strategies require additional cycle time on the instrument and therefore reduce the number of spectra that can be measured in the same amount of time. Additional informatics is often required to make most efficient use of the additional information provided by these spectra as well. This work presents a comparison of several commonly used mass spectrometry methods for the study of phosphopeptide-enriched samples: an MS(2)-only method, a Multistage Activation method, and an MS(2)/MS(3) data-dependent neutral loss method. Several strategies for dealing effectively with the resulting MS(3) data in the latter approach are also presented and compared. The overall goal is to infer whether any one methodology performs significantly better than another for identifying phosphopeptides. On data presented here, the Multistage Activation methodology is demonstrated to perform optimally and does not result in significant loss of unique peptide identifications.

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