Autoren:	Kwiatkowski, Marcel Hotze, Madlen Schumacher, Julia Asif, Abdul R. Miguel, Jose Pittol, Ramos Brenig, Bertram Ramljak, Sanja Zischler, Hans Herlyn, Holger
Titel:	Protein speciation is likely to increase the chance of proteins to be determined in 2-DE/MS
Online-Publikationsdatum:	7-Feb-2023
Erscheinungsdatum:	2022
Sprache des Dokuments:	Englisch
Zusammenfassung/Abstract:	Multiple spotting due to protein speciation might increase a protein's chance of being captured in a random selection of 2-DE spots. We tested this expectation in new (PXD015649) and previously published 2-DE/MS data of porcine and human tissues. For comparison, we included bottom-up proteomics studies (BU-LC/MS) of corresponding biological materials. Analyses of altogether ten datasets proposed that amino acid modification fosters multispotting in 2-DE. Thus, the number of 2-DE spots containing a particular protein more tightly associated with a peptide diversity measure accounting for amino acid modification than with an alternative one disregarding it. Furthermore, every 11th amino acid was a post-translational modification candidate site in 2-DE/MS proteins, whereas in BU-LC/MS proteins this was merely the case in every 21st amino acid. Alternative splicing might contribute to multispotting, since genes encoding 2-DE/MS proteins were found to have on average about 0.3 more transcript variants than their counterparts from BU-LC/MS studies. Correspondingly, resolution completeness as estimated from the representation of transcript variant-rich genes was higher in 2-DE/MS than BU-LC/MS datasets. These findings suggest that the ability to resolve proteomes down to protein species can lead to enrichment of multispotting proteins in 2-DE/MS. Low sensitivity of stains and MS instruments appears to enhance this effect.
DDC-Sachgruppe:	300 Sozialwissenschaften 300 Social sciences
Veröffentlichende Institution:	Johannes Gutenberg-Universität Mainz
Organisationseinheit:	FB 02 Sozialwiss., Medien u. Sport
Veröffentlichungsort:	Mainz
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-8767
Version:	Published version
Publikationstyp:	Zeitschriftenaufsatz
Weitere Angaben zur Dokumentart:	Scientific article
Nutzungsrechte:	CC BY-NC
Informationen zu den Nutzungsrechten:	https://creativecommons.org/licenses/by-nc/4.0/
Zeitschrift:	Electrophoresis 43 11
Seitenzahl oder Artikelnummer:	1203 1214
Verlag:	John Wiley & Sons, Ltd
Verlagsort:	Weinheim
Erscheinungsdatum:	2022
ISSN:	1522-2683
DOI der Originalveröffentlichung:	10.1002/elps.202000393
Enthalten in den Sammlungen:	DFG-491381577-H