LaCyTools: A Targeted Liquid Chromatography-Mass Spectrometry Data Processing Package for Relative Quantitation of Glycopeptides.
Bottom-up glycoproteomics by liquid chromatography-mass spectrometry (LC-MS) is an established approach for assessing glycosylation in a protein- and site-specific manner. Consequently, tools are needed to automatically align, calibrate, and integrate LC-MS glycoproteomics data. We developed a modular software package designed to tackle the individual aspects of an LC-MS experiment, called LaCyTools. Targeted alignment is performed using user defined m/z and retention time (tr) combinations. Subsequently, sum spectra are created for each user defined analyte group. Quantitation is performed on the sum spectra, where each user defined analyte can have its own tr, minimum, and maximum charge states. Consequently, LaCyTools deals with multiple charge states, which gives an output per charge state if desired, and offers various analyte and spectra quality criteria. We compared throughput and performance of LaCyTools to combinations of available tools that deal with individual processing steps. LaCyTools yielded relative quantitation of equal precision (relative standard deviation <0.5%) and higher trueness due to the use of MS peak area instead of MS peak intensity. In conclusion, LaCyTools is an accurate automated data processing tool for high-throughput analysis of LC-MS glycoproteomics data. Released under the Apache 2.0 license, it is freely available on GitHub ( https://github.com/Tarskin/LaCyTools ).
Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300RC Leiden, The Netherlands.
Bas C
BC
Jansen
Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300RC Leiden, The Netherlands.
David
D
Falck
Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300RC Leiden, The Netherlands.
Noortje
N
de Haan
Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300RC Leiden, The Netherlands.
Agnes L
AL
Hipgrave Ederveen
Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry, University of Zagreb , A. Kovačića 1, HR10000 Zagreb, Croatia.
Genadij
G
Razdorov
Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry, University of Zagreb , A. Kovačića 1, HR10000 Zagreb, Croatia.
Gordan
G
Lauc
Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300RC Leiden, The Netherlands.
Manfred
M
Wuhrer
D002853:Chromatography, Liquid; D001330:Electronic Data Processing / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis*; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods* / Q000592:standards; D012984:Software
D006020:Glycopeptides
alpha-1-antitrypsin; bioinformatics; glycoproteomics; immunoglobulin G; liquid chromatography; mass spectrometry; profiling; quality control; relative quantitation