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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
dataset
posted on 2021-10-04, 21:10 authored by Juan J. Calvete, Davinia Pla, Johannes Els, Salvador Carranza, Maik Damm, Benjamin-Florian Hempel, Elisa B. O. John, Daniel Petras, Paul Heiss, Ayse Nalbantsoy, Bayram Göçmen, Roderich D. Süssmuth, Francisco Calderón-Celis, Alicia Jiménez Nosti, Jorge Ruiz EncinarWe report a novel hybrid, molecular
and elemental mass spectrometry
(MS) setup for the absolute quantification of snake venom proteomes
shown here for two desert black cobra species within the genus Walterinnesia, Walterinnesia aegyptia and Walterinnesia morgani. The experimental
design includes the decomplexation of the venom samples by reverse-phase
chromatography independently coupled to four mass spectrometry systems:
the combined bottom-up and top-down molecular MS for protein identification
and 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 quadrupole
time-of-flight mass spectrometry (ESI-QToF MS). This allows to continuously
record the absolute sulfur concentration throughout the chromatogram
and assign it to the parent venom proteins separated in the RP-μHPLC-ESI-QToF
parallel run via mass profiling. The results provide a locus-resolved
and quantitative insight into the three desert black cobra venom proteome
samples. They also validate the units of measure of our snake venomics
strategy for the relative quantification of snake venom proteomes
as % of total venom peptide bonds as a proxy for the % by weight of
the venom toxins/toxin families. In a more general context, our work
may pave the way for broader applications of hybrid elemental/molecular
MS setups in diverse areas of proteomics.
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work may pavephase microbore highperformance liquid chromatographinductively coupled plasmaflight mass spectrometryexperimental design includesdesert black cobrassnake venomics strategyelemental mass spectrometryqtof ms ).>, walterinnesia aegyptiasnake venom proteomesmolecular ms setupswalterinnesia aegyptiavenomics characterizationhybrid elementalvenom toxinsvenom sampleswalterinnesia morganiwalterinnesia molecular mstwo speciestoxin familiesresults providerelative quantificationquantitative insightprotein identificationnovel hybridgeneral contextdiverse areascontinuously recordcombined molecularcombined bottombroader applicationsalso validateabsolute quantification