Combined Molecular and Elemental Mass Spectrometry Approaches for Absolute Quantification of Proteomes: Application to the Venomics Characterization of the Two Species of Desert Black Cobras, Walterinnesia aegyptia and Walterinnesia morgani.

Content Provider	Europe PMC
Author	Calvete, Juan J. Pla, Davinia Els, Johannes Carranza, Salvador Damm, Maik Hempel, Benjamin-Florian John, Elisa B. O. Petras, Daniel Heiss, Paul Nalbantsoy, Ayse Göçmen, Bayram Süssmuth, Roderich D. Calderón-Celis, Francisco Nosti, Alicia Jiménez Encinar, Jorge Ruiz
Copyright Year	2021
Abstract	We report a novel hybrid, molecularand elemental mass spectrometry(MS) setup for the absolute quantification of snake venom proteomesshown here for two desert black cobra species within the genus Walterinnesia, Walterinnesia aegyptia and Walterinnesia morgani. The experimentaldesign includes the decomplexation of the venom samples by reverse-phasechromatography independently coupled to four mass spectrometry systems:the combined bottom-up and top-down molecular MS for protein identificationand a parallel reverse-phase microbore high-performance liquid chromatograph(RP-μHPLC) on-line to inductively coupled plasma (ICP-MS/MS)elemental mass spectrometry and electrospray ionization quadrupoletime-of-flight mass spectrometry (ESI-QToF MS). This allows to continuouslyrecord the absolute sulfur concentration throughout the chromatogramand assign it to the parent venom proteins separated in the RP-μHPLC-ESI-QToFparallel run via mass profiling. The results provide a locus-resolvedand quantitative insight into the three desert black cobra venom proteomesamples. They also validate the units of measure of our snake venomicsstrategy for the relative quantification of snake venom proteomesas % of total venom peptide bonds as a proxy for the % by weight ofthe venom toxins/toxin families. In a more general context, our workmay pave the way for broader applications of hybrid elemental/molecularMS setups in diverse areas of proteomics.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC8576837&blobtype=pdf
ISSN	15353893
Journal	Journal of Proteome Research [J Proteome Res]
Volume Number	20
DOI	10.1021/acs.jproteome.1c00608
PubMed Central reference number	PMC8576837
Issue Number	11
PubMed reference number	34606723
e-ISSN	15353907
Language	English
Publisher	American Chemical Society
Publisher Date	2021-10-04
Access Restriction	Open
Rights License	Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). © 2021 American Chemical Society
Subject Keyword	snake venomics combined top-down and bottom-upvenomics hybrid elemental and molecular mass spectrometry absolutequantification of venom proteome desert black cobra Walterinnesia aegyptia Walterinnesia morgani
Content Type	Text
Resource Type	Article
Subject	Chemistry Biochemistry