The high-throughput production of membrane proteins
| Autor: | James M. Birch, A. Quigley |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
High-throughput screening
Cell Computational biology high-throughput screening General Biochemistry Genetics and Molecular Biology Biochemical Techniques & Resources Structural Biology transmembrane proteins protein purification Protein purification medicine Humans protein expression Molecular Biology Crystallography Expression vector Chemistry Cryoelectron Microscopy Membrane Proteins Transmembrane protein medicine.anatomical_structure Structural biology Membrane protein Perspective Cell Membranes Excitation & Transport General Agricultural and Biological Sciences Function (biology) |
| Zdroj: | Emerging Topics in Life Sciences |
| ISSN: | 2397-8562 2397-8554 |
| DOI: | 10.1042/etls20210196 |
| Popis: | Membrane proteins, found at the junctions between the outside world and the inner workings of the cell, play important roles in human disease and are used as biosensors. More than half of all therapeutics directly affect membrane protein function while nanopores enable DNA sequencing. The structural and functional characterisation of membrane proteins is therefore crucial. However, low levels of naturally abundant protein and the hydrophobic nature of membrane proteins makes production difficult. To maximise success, high-throughput strategies were developed that rely upon simple screens to identify successful constructs and rapidly exclude those unlikely to work. Parameters that affect production such as expression host, membrane protein origin, expression vector, fusion-tags, encapsulation reagent and solvent composition are screened in parallel. In this way, constructs with divergent requirements can be produced for a variety of structural applications. As structural techniques advance, sample requirements will change. Single-particle cryo-electron microscopy requires less protein than crystallography and as cryo-electron tomography and time-resolved serial crystallography are developed new sample production requirements will evolve. Here we discuss different methods used for the high-throughput production of membrane proteins for structural biology. |
| Databáze: | OpenAIRE |
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