LTQ-iQuant: A freely available software pipeline for automated and accurate protein quantification of isobaric tagged peptide data from LTQ instruments.

LTQ-iQuant: A freely available software pipeline for automated and accurate protein quantification of isobaric tagged peptide data from LTQ instruments.
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DOI:
10.1002/pmic.201000189
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发表时间:
2010-10
期刊:
影响因子:
3.4
通讯作者:
Griffin, Timothy J.
Griffin, Timothy J.
中科院分区:
生物学3区
文献类型:
--
作者:
Onsongo, Getiria;Stone, Matthew D.;Van Riper, Susan K.;Chilton, John;Wu, Baolin;Higgins, LeeAnn;Lund, Troy C.;Carlis, John V.;Griffin, Timothy J.

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Pulsed Q dissociation enables combining LTQ ion trap instruments with isobaric peptide tagging. Unfortunately, this combination lacks a technique which accurately reports protein abundance ratios and is implemented in a freely-available, flexible software pipeline. We developed and implemented a technique assigning collective reporter ion intensity-based weights to each peptide abundance ratio and calculating a protein’s weighted average abundance ratio and P value. Using an iTRAQ-labeled standard mixture, we compared our technique’s performance to the commercial software Mascot, finding that it performed better than Mascot’s non-weighted averaging and median peptide ratio techniques, and equal to its weighted averaging technique. We also compared performance of the LTQ-Orbitrap plus our technique to 4800 MALDI TOF/TOF plus Protein Pilot, by analyzing an iTRAQ-labeled stem cell lysate. We found highly correlated protein abundance ratios, indicating that the LTQ-Orbitrap plus our technique yields results comparable to the current standard. We implemented our technique in a freely available, automated software pipeline, called LTQ-iQuant, which: is mzXML-compatible; supports iTRAQ 4-plex and 8-plex LTQ data; and can be modified for and have weights trained to a user’s LTQ and other isobaric peptide tagging methods. LTQ-iQuant should make LTQ instruments and isobaric peptide tagging accessible to more proteomics researchers.
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