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 .

Autor:	Calvete JJ; Laboratorio de Venómica Evolutiva y Traslational, Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Jaume Roig 11, 46010 Valencia, Spain., Pla D; Laboratorio de Venómica Evolutiva y Traslational, Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Jaume Roig 11, 46010 Valencia, Spain., Els J; Environment and Protected Areas Authority, 82828 Sharjah, United Arab Emirates., Carranza S; Institute of Evolutionary Biology, CSIC-Universitat Pompeu Fabra, 08003 Barcelona, Spain., Damm M; Department of Chemistry, Technische Universität Berlin, 10623 Berlin, Germany., Hempel BF; Department of Chemistry, Technische Universität Berlin, 10623 Berlin, Germany.; BIH Center for Regenerative Therapies BCRT, Charité-Universitätsmedizin Berlin, 13353 Berlin, Germany., John EBO; Center of Biotechnology, Universidade Federal do Rio Grande do Sul, CEP 91501-970 Porto Alegre, RS, Brazil., Petras D; CMFI Cluster of Excellence, Interfaculty Institute of Microbiology and Medicine, University of Túbingen, 72076 Tübingen, Germany., Heiss P; Department of Chemistry, Technische Universität Berlin, 10623 Berlin, Germany., Nalbantsoy A; Department of Bioengineering, Faculty of Engineering, Ege University, 35100 Bornova, Izmir, Turkey., Göçmen B; Zoology Section, Department of Biology, Faculty of Science, Ege University, 35100 Bornova, Izmir, Turkey., Süssmuth RD; Department of Chemistry, Technische Universität Berlin, 10623 Berlin, Germany., Calderón-Celis F; Department of Physical and Analytical Chemistry, University of Oviedo, 33006 Oviedo, Spain., Nosti AJ; Department of Physical and Analytical Chemistry, University of Oviedo, 33006 Oviedo, Spain., Encinar JR; Department of Physical and Analytical Chemistry, University of Oviedo, 33006 Oviedo, Spain.
Jazyk:	angličtina
Zdroj:	Journal of proteome research [J Proteome Res] 2021 Nov 05; Vol. 20 (11), pp. 5064-5078. Date of Electronic Publication: 2021 Oct 04.
DOI:	10.1021/acs.jproteome.1c00608
Abstrakt:	We 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.
Databáze:	MEDLINE
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