Biochemical and proteomic profiling reveals potent proteolytic activity in the venom of Erythrolamprus melanotus (Shaw's dark ground snake, Xenodontini)

Author(s)

Torres-Bonilla, Kristian A. ; Bayona-Serrano, Juan D. ; Saenz-Suarez, Paula A. ; Zambrano, Duvan F. ; Bernal-Bautista, Manuel H. ; Junqueira-de-Azevedo, Inacio L. M. ; Hyslop, Stephen

• Document type: Journal article
• Source: Biochimie; v. 235, p. 12-pg., 2025-08-01.

Abstract

We describe here the Duvernoy's venom gland organization and biochemical and proteomic analyses of venom from the Colombian colubrid snake Erythrolamprus melanotus. Histological analysis indicated a serous venom gland distinct from the mucous supralabial gland. SDS-PAGE of the venom revealed >15 bands (similar to 15 kDa-similar to 200 kDa) and gelatin zymography revealed potent proteolytic activity; zymographic activity towards casein was considerably less. RP-HPLC yielded 16 peaks, several of which contained proteases, as assessed by gelatin zymography. Enzymatic assays using azocasein, azocoll and hide powder azure confirmed the proteolytic activity. The venom selectively degraded the A alpha-chain of fibrinogen without affecting the B beta and gamma-chains (assessed by SDS-PAGE). All proteolytic activity was inhibited by EDTA (metalloproteinase inhibitor) but not by AEBSF (serine proteinase inhibitor). The venom had no L-amino acid oxidase, esterase, and PLA(2) activities. Proteomic analysis indicated the presence primarily of snake venom metalloproteinases (SVMP), snake venom matrix metalloproteinases (svMMP) and cysteine-rich secretory proteins (CRiSP). SVMP and svMMP were the major components, indicating that this venom is rich in proteases, in agreement with the findings for zymography and enzymatic assays. This study provides the first characterization of Colombian E. melanotus venom and highlights its rich content of metalloproteinases and potent proteolytic activity. (c) 2025 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights are reserved, including those for text and data mining, AI training, and similar technologies. (AU)

• FAPESP's process: 24/14508-0 - Technical support for CeTICS genomics projects: sequencing and assembly of snake genomes
• Grantee: Juan David Bayona Serrano
• Support Opportunities: Scholarships in Brazil - Technical Training Program - Technical Training
• FAPESP's process: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling
• Grantee: Hugo Aguirre Armelin
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 16/50127-5 - Dimensions US-BIOTA São Paulo: scales of biodiversity: integrated studies of snake venom evolution and function across multiple levels of diversity
• Grantee: Inácio de Loiola Meirelles Junqueira de Azevedo
• Support Opportunities: BIOTA-FAPESP Program - Thematic Grants
• FAPESP's process: 17/24498-9 - Venomics analyses of Dipsadinae snakes focusing on adaptations to ophiophagy
• Grantee: Juan David Bayona Serrano
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)