advertisement

Yasset perezriverol csi2011

60 %
40 %
advertisement
Information about Yasset perezriverol csi2011

Published on March 5, 2014

Author: ypriverol

Source: slideshare.net

advertisement

In silico analysis of accurate proteomics, complemented by selective isolation of peptides. Yasset Perez-Riverol yperez@ebi.ac.uk yasset.perez@biocomp.cigb.edu.cu Aniel Sanchez Puentes aniel.sanchez@cigb.edu.cu EBI is an Outstation of the European Molecular Biology Laboratory.

ABSTRACT Protein identification by mass spectrometry is mainly based on MS/MS spectra and the accuracy of molecular mass determination. However, the high complexity and dynamic ranges for any species of proteomic samples, surpasses the separation capacity and detection power of the most advanced multidimensional liquid chromatographs and mass spectrometers. Only a tiny portion of signals is selected for MS/MS experiments and a still considerable number of them do not provide reliable peptide identification. The approach is based on mass accuracy, isoelectric point (pI), retention time (tR) and N-terminal amino acid determination as protein identification criteria regardless of high quality MS/MS spectra. When the methodology was combined with the selective isolation methods, the number of unique peptides and identified proteins increases. Finally, to demonstrate the feasibility of the methodology, an OFFGEL-LC-MS/MS experiment was also implemented. Our results show that using the information provided by these features and selective isolation methods we could found the 93% of the high confidence protein identified by MS/MS with false-positive rate lower than 5%.

Drosophila cell RH0 ~~~~ K RH1 ~~~~R (0) Abs at 215 nm ~~H~~K RH2 ~~H~~R (1+) (2+, 3+) 20 10 30 40 764.974 100 b % 1 292.163 RPEGENASYHLAYDKDR389 373 764.320 719.299 0 827.868 100 b % 1 251.126 0 b 100 110.025 1 292.138 % 0 y n 828.373 DSSIVTHDNDIFR233 221 828.889 702.367 1 RPEGENASYHLAYDK389 373 1021.131 1238.504 1450.501 1637.761 1140.3881333.778 1577.739 1762.484 MS/MS spectra were interpreted by the X! Tandem software using the Flybase sequence database. The database search results were validated using PeptideProphet. This work analyzed only the four isoelectric focusing fractions with the lowest pI having the best agreement between the theoretical and experimental values, according to previous. In addition, these fractions cover 50% of the identified peptides. Also, we used only highly reliable peptide identifications, filtering out those with a PeptideProphet probability lower than 0.97 (FDR = 0.01) or with posttranslational modifications. For experimental t R analysis the acceptance error was set at 748.42 s, and mass tolerance was set at 10 ppm.

Proteomic Research: N-Term Identification Anal Chem. 2010 Oct 15;82(20):8492-501.

Create a Insilico tryptic peptide database. Annotate peptides with theoretical rt, pI, mass, MW, N-Term Annotate experimental identified sequences from PeptideProphet output with probability more than 0.97. (rt, pI, N-term, MW, sequence) Search precursor masses of Experimental sequences on Insilco Database. [Match only one sequence] [Not match any sequence] Peptides out of the ppm range [Match with more than one sequence] Search in the input theoretical List of sequence the peptide By current property (pI, rt, MW, N-term) [Not match any sequence] [Match with more than one sequence] Peptides out of the error range for property [Match only one sequence] Compare with MS/MS sequence result [Not Match Insilco sequence with MS/MS sequence] Annotate as a False Positive Identification. [Match Insilco sequence with MS/MS sequence] Annotate as Peptide Identification A tree-based algorithm to identify unique peptides in the experimental set was constructed in a similar fashion to the one designed for theoretical analysis. The final list of unique peptides was validated by using the sequence predicted from PeptideProphet. In cases where the PeptideProphet sequences and the sequences identified by our approach did not match, the identifications achieved by our algorithm were considered as false positive identification.

CONCLUSION The use of the information provided by some analytical tools could help to offset the information contained in the sequence of peptides, but it is more efficient when a prokaryote proteome is analyzed. Some drawbacks associated to precision (accuracy) that can be predicted are that the variables used may hinder the accurate mass proteomics analysis with the identification of false positive hints. The inclusion of some types of peptides and the reduction of complexity allows increasing the percent of unique peptides compared to normal analysis. The combination of several selective methods (RH0, RH1, and RH2) in the same sample could increase the percent of proteins with unique peptides. The theoretical analysis described in this paper does not exclude the possibility of combining it with the MS/MS information obtained in any proteomic experiment.

Add a comment

Related pages

Yasset Perez-Riverol | LinkedIn

View Yasset Perez-Riverol’s professional profile on LinkedIn. LinkedIn is the world's largest business network, helping professionals like Yasset Perez ...
Read more

Yasset Perez-Riverol | LinkedIn

View Yasset Perez-Riverol’s professional profile on LinkedIn. LinkedIn is the world's largest business network, helping professionals like Yasset Perez ...
Read more

Yasset Perez-Riverol | LinkedIn

View Yasset Perez-Riverol’s professional profile on LinkedIn. LinkedIn is the world's largest business network, helping professionals like Yasset Perez ...
Read more

Yasset Perez-Riverol | LinkedIn

Ver el perfil profesional de Yasset Perez-Riverol en LinkedIn. LinkedIn es la red de negocios más grande del mundo que ayuda a profesionales como Yasset ...
Read more

Yasset Perez-Riverol | LinkedIn

Ver el perfil profesional de Yasset Perez-Riverol en LinkedIn. LinkedIn es la red de negocios más grande del mundo que ayuda a profesionales como Yasset ...
Read more

Yasset Perez-Riverol | LinkedIn

Yasset Perez-Riverols berufliches Profil anzeigen LinkedIn ist das weltweit größte berufliche Netzwerk, das Fach- und Führungskräften wie Yasset Perez- ...
Read more

Yasset Perez-Riverol | LinkedIn

Yasset Perez-Riverol. Senior Software Engineer. Locatie Hinxton, Cambridgeshire, Verenigd Koninkrijk Bedrijfstak Onderzoek en wetenschap
Read more

Yasset Perez-Riverol | LinkedIn

Ver el perfil profesional de Yasset Perez-Riverol en LinkedIn. LinkedIn es la red de negocios más grande del mundo que ayuda a profesionales como Yasset ...
Read more

Yasset Perez-Riverol - HubSlide

HubSlide is the easiest way to upload & share PowerPoint & PDF presentations online; embed in Blogs, Slideshows, Transcript & more
Read more