Article: Proteogenomics of Malignant Melanoma Cell Lines: The Effect of Stringency of Exome Data Filtering on Variant Peptide Identification in Shotgun Proteomics
Journal of proteome research. 2018 Apr. 05, v. 17, no. 5
2018
Abstract: The identification of genetically encoded variants at the proteome level is an important problem in cancer proteogenomics. The generation of customized protein databases from DNA or RNA sequencing data is a crucial stage of the identification workflow. ... ...
| Abstract | The identification of genetically encoded variants at the proteome level is an important problem in cancer proteogenomics. The generation of customized protein databases from DNA or RNA sequencing data is a crucial stage of the identification workflow. Genomic data filtering applied at this stage may significantly modify variant search results, yet its effect is generally left out of the scope of proteogenomic studies. In this work, we focused on this impact using data of exome sequencing and LC–MS/MS analyses of six replicates for eight melanoma cell lines processed by a proteogenomics workflow. The main objectives were identifying variant peptides and revealing the role of the genomic data filtering in the variant identification. A series of six confidence thresholds for single nucleotide polymorphisms and indels from the exome data were applied to generate customized sequence databases of different stringency. In the searches against unfiltered databases, between 100 and 160 variant peptides were identified for each of the cell lines using X!Tandem and MS-GF+ search engines. The recovery rate for variant peptides was ∼1%, which is approximately three times lower than that of the wild-type peptides. Using unfiltered genomic databases for variant searches resulted in higher sensitivity and selectivity of the proteogenomic workflow and positively affected the ability to distinguish the cell lines based on variant peptide signatures. |
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| Keywords | cell lines ; DNA ; genetic databases ; genomics ; liquid chromatography ; melanoma ; peptides ; proteome ; proteomics ; sequence analysis ; single nucleotide polymorphism ; tandem mass spectrometry | |||||
| Language | English | |||||
| Dates of publication | 2018-0405 | |||||
| Size | p. 1801-1811. | |||||
| Publishing place | American Chemical Society | |||||
| Document type | Article | |||||
| ZDB-ID | 2078618-9 | |||||
| ISSN | 1535-3907 ; 1535-3893 | |||||
| ISSN (online) | 1535-3907 | |||||
| ISSN | 1535-3893 | |||||
| DOI | 10.1021/acs.jproteome.7b00841 | |||||
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| Database | NAL-Catalogue (AGRICOLA) |
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