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Mass Spectrometry Differentiation between Rana arvalis Populations Based on Their Skin Peptidome Composition
journal contributionposted on 2022-07-13, 03:13 authored by Tatiana Yu. Samgina, Irina D. Vasileva, Polonca Trebse, Gregor Torkar, Alexey K. Surin, Zhaowei Meng, Roman A. Zubarev, Albert T. Lebedev
Skin secretion of amphibians often represents the only weapon of these species against pathogens and predators. Peptides constitute the major portion of active molecules of that weapon and may be treated as potential pharmaceuticals for future generations. The first step of their efficient use involves establishing of their primary structure, i.e., sequencing. De novo sequencing by means of mass spectrometry was applied to Rana arvalis species, collected in the spring 2021 in Central Slovenia (vicinity of Ljubljana). HPLC-ESI-HRMS/MS with Orbitrap instruments was used to establish the skin peptidome of these species and compare it with the earlier identified skin peptidome of the Moscow population of Rana arvalis. Application of CID, HCD, ETD, and EThcD enabled detecting and sequencing 18 peptides; five of them were novel and may be treated as possible biomarkers of the Ljubljana population of Rana arvalis. Interestingly, representatives of two peptide families (temporins and brevinins 2) were not found in the Moscow population. MS3 modes, first of all EThcD, demonstrated their great potential in the de novo sequencing, including extraction of the sequence information from the intact peptides with disulfide cycle (rana box) in their structure and differentiation of isomeric Leu/Ile residues. Thus, all six isomeric residues were reliably distinguished in the novel melittin-related peptide AK-23-1. In addition, another post-translational modification dealing with carbonylation of the N-terminal Gly of novel temporin AVa was established using the MS3 mode. The obtained results demonstrate the efficiency of the use of MS3 tools in proteomics/peptidomics.
two peptide familiestranslational modification dealingrelated peptide akobtained results demonstratede novo amphibians often representsrana arvalis rana arvalis .e ., sequencingsix isomeric residuesnovel temporin avaethcd enabled detecting3 supsequencing 18 peptidesmass spectrometry differentiationmass spectrometryrana boxisomeric leuile residuespeptides constituteintact peptidesterminal glyspring 2021skin peptidomesequence informationreliably distinguishedpotential pharmaceuticalspossible biomarkerspopulations basedorbitrap instrumentsnovel melittinmoscow populationmajor portionljubljana populationljubljana ).including extractiongreat potentialfuture generationsestablished usingdisulfide cyclecentral sloveniabrevinins 2another postactive molecules