Synthesizing Membrane Proteins Using In Vitro Methodology (ANL-IN-06-099)
Scientists at Argonne National Laboratory have created an in vitro, cell-free system and method for producing several types of protein: membrane proteins, membrane-associated proteins, and soluble proteins.
With advances that can be gleaned from the study of high quality samples of this type, this method is expected to drive advances in membrane protein structural biology and deepen our approaches for characterizing biological activity as cellular interfaces.Description
In most organisms, cell membranes are the vital structures that serve as the interface between an organism and its environment, enabling the creation of compartments where proteins carry out the cell’s basic functions. Proteins in these membranes carry out the essential functions of the cell, such as uptake of nutrients, excretion of wastes, energy generation, and signal transduction. The functions performed by membrane proteins are extremely important for all organisms. Previously, researchers studying these proteins needed to replicate them within cells: a complex, time-consuming process.
Despite the fact that they represent approximately 30% of every genome and comprise more than 60% of all drug targets, only about 100 unique membrane protein structures have been determined to date (compared to about 10,000 unique structures in soluble protein families). One reason for this relatively low number of unique membrane protein structures is that it is difficult to isolate membrane proteins using conventional methods. Also, once isolated, purification is highly protein-specific, is not adaptable to high-throughput methodologies, and rarely yields the amounts of pure membrane proteins needed for extensive biochemical studies and crystallization trials.
The in vitro method is capable of producing membrane proteins, membrane-associated proteins, and soluble proteins. This methodology promises to become an important tool for deepening scientists’ understanding of life and driving advances in molecular biology.Benefits
Without the need for a host cell, scientists can synthesize proteins—even proteins that are toxic or fragile in composition—more quickly and more easily than by conventional methodologies.Applications and Industries
- Medical research
- Pharmaceuticals and the development of new drugs and therapies
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Cell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins
The invention provides an in vitro method for producing proteins, membrane proteins, membrane-associated proteins, and soluble proteins that interact with membrane-associated proteins for assembly into an oligomeric complex or that require association with a membrane for proper folding. The method comprises, supplying intracytoplasmic membranes from organisms; modifying protein composition of intracytoplasmic membranes from organism by modifying DNA to delete genes encoding functions of the organism not associated with the formation of the intracytoplasmic membranes; generating appropriate DNA or RNA templates that encode the target protein; and mixing the intracytoplasmic membranes with the template and a transcription/translation-competent cellular extract to cause simultaneous production of the membrane proteins and encapsulation of the membrane proteins within the intracytoplasmic membranes.
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