Distinct bothrojaracin isoforms produced by individual jararaca (Bothrops jararaca) snakes

Area of Science:

• Biochemistry
• Venomics
• Molecular Biology

Background:

• Bothrojaracin is a potent thrombin inhibitor found in Bothrops jararaca venom.
• Previous studies identified multiple molecular variants (isoforms) in pooled venom.
• The production of single versus multiple isoforms by individual snakes remained uncharacterized.

Purpose of the Study:

• To investigate whether individual Bothrops jararaca snakes produce a single type or multiple isoforms of bothrojaracin.
• To analyze the molecular heterogeneity of bothrojaracin within individual snake venoms.

Main Methods:

• Individual venom collection from six adult Bothrops jararaca snakes.
• Purification of bothrojaracin using alpha-thrombin affinity chromatography and Superose 12 HR gel filtration.
• Analysis of purified proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and amino-terminal sequencing.

Main Results:

• Bothrojaracin was detected in all individual venoms, with high activity in three samples.
• Purified proteins from high-activity venoms showed single bands (27,000 Da) on SDS-PAGE with homologous amino-terminal sequences.
• Venoms with lower activity contained bothrojaracin and inactive proteins.
• Sequence analysis revealed distinct isoforms within the venom of individual snakes.

Conclusions:

• Individual Bothrops jararaca snakes can produce multiple, distinct isoforms of bothrojaracin.
• Sequence differences in bothrojaracin highlight molecular diversity within a single snake's venom.
• This finding contributes to understanding the complexity of snake venom composition and evolution.