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A Rapid and Reliable Method for Total Protein Extraction from Succulent Plants for Proteomic Analysis
Fernando Lledías1, Felipe Hernández2, Viridiana Rivas1
1Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad # 2001, Col. Chamilpa C.P., 62210, Cuernavaca, Morelos, Mexico.
The Protein Journal
|May 13, 2017
Summary
A new, rapid protocol effectively extracts proteins from challenging succulent plants, enabling molecular studies. This method overcomes issues caused by complex compounds, yielding high-quality protein patterns for SDS/PAGE and 2DE analysis.
Area of Science:
- Plant molecular biology
- Biochemistry
- Proteomics
Background:
- Succulent plants possess unique morphological and molecular adaptations for xeric environments.
- These adaptations involve compounds like polysaccharides, pigments, waxes, and terpenoids that interfere with protein analysis.
- Existing protein extraction protocols are inadequate for these plants, hindering molecular-level research.
Purpose of the Study:
- To develop a rapid, reliable, and broadly applicable protocol for protein extraction from succulent plants.
- To enable molecular and proteomic studies in plant groups with challenging biochemical compositions.
Main Methods:
- A novel protein extraction method utilizing differential solubility in acetone and pH-altered solutions.
- Application of the protocol to diverse succulent species from Agavoideae, Nolinoideae, and Cactaceae families.
- Adaptation of the protocol for two-dimensional electrophoresis (2DE) for proteomic analysis.
Main Results:
- Successfully obtained well-resolved SDS/PAGE protein patterns from various succulent plant extracts.
- Demonstrated the protocol's effectiveness across different subfamilies and families of succulent plants.
- Identified Furostanol glycoside 26-O-β-glucosidase in Agave attenuata using 2DE and mass spectrometry.
Conclusions:
- The developed protocol significantly overcomes challenges in protein extraction from succulent plants.
- This method facilitates molecular and proteomic investigations in these ecologically important plants.
- The protocol supports reproducible protein analysis, including advanced techniques like 2DE and mass spectrometry.
