Abstract
Mass spectrometry (MS)-based proteomics, which uses high-resolution hybrid mass spectrometers such as the quadrupole-orbitrap mass spectrometer, can yield tens of thousands of tandem mass (MS/MS) spectra of high resolution during a routine bottom-up experiment. Despite being a fundamental and key step in MS-based proteomics, the accurate determination and assignment of precursor monoisotopic masses to the MS/MS spectra remains difficult. The difficulties stem from imperfect isotopic envelopes of precursor ions, inaccurate charge states for precursor ions, and cofragmentation. We describe a composite method of utilizing MS data to assign accurate monoisotopic masses to MS/MS spectra, including those subject to cofragmentation. The method, "multiplexed post-experiment monoisotopic mass refinement" (mPE-MMR), consists of the following: multiplexing of precursor masses to assign multiple monoisotopic masses of cofragmented peptides to the corresponding multiplexed MS/MS spectra, multiplexing of charge states to assign correct charges to the precursor ions of MS/ MS spectra with no charge information, and mass correction for inaccurate monoisotopic peak picking. When combined with MS-GF+, a database search algorithm based on fragment mass difference, mPE-MMR effectively increases both sensitivity and accuracy in peptide identification from complex high-throughput proteomics data compared to conventional methods.
| Original language | English |
|---|---|
| Pages (from-to) | 1244-1253 |
| Number of pages | 10 |
| Journal | Analytical Chemistry |
| Volume | 89 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2017 Jan 1 |
Fingerprint
ASJC Scopus subject areas
- Analytical Chemistry
Cite this
Multiplexed post-experimental monoisotopic mass refinement (mPE-MMR) to increase sensitivity and accuracy in peptide identifications from tandem mass spectra of cofragmentation. / Madar, Inamul Hasan; Ko, Seung Ik; Kim, Hokeun; Mun, Dong Gi; Kim, Sangtae; Smith, Richard D.; Lee, Sang-Won.
In: Analytical Chemistry, Vol. 89, No. 2, 01.01.2017, p. 1244-1253.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Multiplexed post-experimental monoisotopic mass refinement (mPE-MMR) to increase sensitivity and accuracy in peptide identifications from tandem mass spectra of cofragmentation
AU - Madar, Inamul Hasan
AU - Ko, Seung Ik
AU - Kim, Hokeun
AU - Mun, Dong Gi
AU - Kim, Sangtae
AU - Smith, Richard D.
AU - Lee, Sang-Won
PY - 2017/1/1
Y1 - 2017/1/1
N2 - Mass spectrometry (MS)-based proteomics, which uses high-resolution hybrid mass spectrometers such as the quadrupole-orbitrap mass spectrometer, can yield tens of thousands of tandem mass (MS/MS) spectra of high resolution during a routine bottom-up experiment. Despite being a fundamental and key step in MS-based proteomics, the accurate determination and assignment of precursor monoisotopic masses to the MS/MS spectra remains difficult. The difficulties stem from imperfect isotopic envelopes of precursor ions, inaccurate charge states for precursor ions, and cofragmentation. We describe a composite method of utilizing MS data to assign accurate monoisotopic masses to MS/MS spectra, including those subject to cofragmentation. The method, "multiplexed post-experiment monoisotopic mass refinement" (mPE-MMR), consists of the following: multiplexing of precursor masses to assign multiple monoisotopic masses of cofragmented peptides to the corresponding multiplexed MS/MS spectra, multiplexing of charge states to assign correct charges to the precursor ions of MS/ MS spectra with no charge information, and mass correction for inaccurate monoisotopic peak picking. When combined with MS-GF+, a database search algorithm based on fragment mass difference, mPE-MMR effectively increases both sensitivity and accuracy in peptide identification from complex high-throughput proteomics data compared to conventional methods.
AB - Mass spectrometry (MS)-based proteomics, which uses high-resolution hybrid mass spectrometers such as the quadrupole-orbitrap mass spectrometer, can yield tens of thousands of tandem mass (MS/MS) spectra of high resolution during a routine bottom-up experiment. Despite being a fundamental and key step in MS-based proteomics, the accurate determination and assignment of precursor monoisotopic masses to the MS/MS spectra remains difficult. The difficulties stem from imperfect isotopic envelopes of precursor ions, inaccurate charge states for precursor ions, and cofragmentation. We describe a composite method of utilizing MS data to assign accurate monoisotopic masses to MS/MS spectra, including those subject to cofragmentation. The method, "multiplexed post-experiment monoisotopic mass refinement" (mPE-MMR), consists of the following: multiplexing of precursor masses to assign multiple monoisotopic masses of cofragmented peptides to the corresponding multiplexed MS/MS spectra, multiplexing of charge states to assign correct charges to the precursor ions of MS/ MS spectra with no charge information, and mass correction for inaccurate monoisotopic peak picking. When combined with MS-GF+, a database search algorithm based on fragment mass difference, mPE-MMR effectively increases both sensitivity and accuracy in peptide identification from complex high-throughput proteomics data compared to conventional methods.
UR - http://www.scopus.com/inward/record.url?scp=85035116865&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85035116865&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.6b03874
DO - 10.1021/acs.analchem.6b03874
M3 - Article
C2 - 27966901
AN - SCOPUS:85035116865
VL - 89
SP - 1244
EP - 1253
JO - Industrial And Engineering Chemistry Analytical Edition
JF - Industrial And Engineering Chemistry Analytical Edition
SN - 0003-2700
IS - 2
ER -