Proteomic and Glycoproteomic Profilings Reveal That Post-translational Modifications of Toxins Contribute to Venom Phenotype in Snakes.
Snake venoms are biological weapon systems composed of secreted proteins and peptides that are used for immobilizing or killing prey. Although post-translational modifications are widely investigated because of their importance in many biological phenomena, we currently still have little understanding of how protein glycosylation impacts the variation and stability of venom proteomes. To address these issues, here we characterized the venom proteomes of seven Bothrops snakes using a shotgun proteomics strategy. Moreover, we compared the electrophoretic profiles of native and deglycosylated venoms and, in order to assess their subproteomes of glycoproteins, we identified the proteins with affinity for three lectins with different saccharide specificities and their putative glycosylation sites. As proteinases are abundant glycosylated toxins, we examined the effect of N-deglycosylation on their catalytic activities and show that the proteinases of the seven venoms were similarly affected by removal of N-glycans. Moreover, we prospected putative glycosylation sites of transcripts of a B. jararaca venom gland data set and detected toxin family related patterns of glycosylation. Based on our global analysis, we report that Bothrops venom proteomes and glycoproteomes contain a core of components that markedly define their composition, which is conserved upon evolution in parallel to other molecular markers that determine their phylogenetic classification.
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.
Débora
D
Andrade-Silva
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.|Instituto de Ciência e Tecnologia, Universidade Federal de São Paulo (ICT-UNIFESP), São José dos Campos 12231-280, Brazil.
André
A
Zelanis
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.
Eduardo S
ES
Kitano
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.
Inácio L M
IL
Junqueira-de-Azevedo
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.
Marcelo S
MS
Reis
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.|Departamento de Ciências Exatas e da Terra, Universidade Federal de São Paulo , Diadema 04021-001, Brazil.
Aline S
AS
Lopes
Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan , São Paulo 05503-000, Brazil.
Solange M T
SM
Serrano
D000818:Animals; D001665:Binding Sites; D017837:Bothrops; D003435:Crotalid Venoms / Q000737:chemistry* / Q000201:enzymology; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D010447:Peptide Hydrolases; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012911:Snakes
D003435:Crotalid Venoms; D006023:Glycoproteins; D020543:Proteome; D010447:Peptide Hydrolases
glycoproteome; lectin-affinity chromatography; mass spectrometry; peptidome; proteome; snake venom; transcriptome