Crotalus durissus terrificus plasma proteome

Description

Little is known about the plasma proteome of snakes, which may provide valuable biomarkers for health monitoring and novel therapeutic applications. This study aimed to characterize the plasma proteome of healthy adult Crotalus durissus terrificus snakes to establish a molecular baseline for physiological assessment and potential diagnostic use.

Steps to reproduce

Plasma proteins were subjected to in-gel digestion according to the protocol described by Shevchenko et al. (2006). The resulting peptides were analyzed by nano-LC-ESI-MS/MS using a MicrQ-TOF III quadrupole time-of-flight mass spectrometer (Bruker Daltonics) interfaced with a Shimadzu LC-20AT system equipped with a binary pump and autosampler. Peptide separation was carried out on a reverse-phase C18 column (4.5 × 100 mm, 1.8 µm) under a linear gradient ranging from 0 to 85% acetonitrile in 0.1% formic acid or TFA for 60 minutes, at a flow rate of 0.2 mL/min. The column was maintained at 25 °C, while the autosampler was kept at 10 °C. Mass spectrometry acquisition was performed in positive ionization mode at 4.5 kV, with source temperature set to 180 °C, nitrogen gas flow at 6 L/min, and a pressure of 0.8 bar, scanning ions within the 100–1000 m/z range. Tandem MS was carried out using nitrogen as the collision gas with energies from 1 to 200 eV. Prior to injection, samples were dissolved in 50% acetonitrile containing 0.1% TFA and introduced at a flow rate of 0.3 mL/min. Instrument calibration was performed both internally and externally with sodium formate, and spectra were processed using Bruker Data Analysis software version 3.3.

Institutions

• Universidade Estadual Paulista Julio de Mesquita Filho Faculdade de Medicina Campus de Botucatu

  SP, Botucatu

Categories

Funders

• National Council for Scientific and Technological Development

  Federal District, Brazil

  Grant ID: 437089/2018-5

Licence