Decision peptide-driven: A free software tool for accurate protein quantification using gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry
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- 作者：Hugo M. Santos ; Miguel Reboiro-Jato ; Daniel Glez-Peñ ; a ; J.D. Nunes-Mir ; a ; Florentino Fdez-Riverola ; R. Carvallo ; J.L. Capelo
- 关键词：Protein quantification ; Gel electrophoresis ; MALDI-TOF-MS ; 18O-labeling ; DPD software
- 刊名：Talanta
- 出版年：2010
- 出版时间：15 September 2010
- 年：2010
- 卷：82
- 期：4
- 页码：1412-1420
- 全文大小：949 K
文摘
The decision peptide-driven tool implements a software application for assisting the user in a protocol for accurate protein quantification based on the following steps: (1) protein separation through gel electrophoresis; (2) in-gel protein digestion; (3) direct and inverse 18O-labeling and (4) matrix assisted laser desorption ionization time of flight mass spectrometry, MALDI analysis. The DPD software compares the MALDI results of the direct and inverse 18O-labeling experiments and quickly identifies those peptides with paralleled loses in different sets of a typical proteomic workflow. Those peptides are used for subsequent accurate protein quantification. The interpretation of the MALDI data from direct and inverse labeling experiments is time-consuming requiring a significant amount of time to do all comparisons manually. The DPD software shortens and simplifies the searching of the peptides that must be used for quantification from a week to just some minutes. To do so, it takes as input several MALDI spectra and aids the researcher in an automatic mode (i) to compare data from direct and inverse 18O-labeling experiments, calculating the corresponding ratios to determine those peptides with paralleled losses throughout different sets of experiments; and (ii) allow to use those peptides as internal standards for subsequent accurate protein quantification using 18O-labeling. In this work the DPD software is presented and explained with the quantification of protein carbonic anhydrase.