1.3 Efficient Strategies for Production of Eukaryotic Proteins

Cumulative experience based on structural genomics of thousands of eukaryotic proteins enables a rational and informed approach to be taken when setting up a new program for any level of throughput. This chapter presents strategies for the production of proteins for structural or biochemical functional studies. Emphasis is placed on delivery of high-purity, stable proteins in a format suitable for crystallization or nuclear magnetic resonance spectroscopy and having defined biochemical function. A critical component relates to design and selection of a polypeptide with the minimal sequence determinants for a particular biochemical activity, in order to facilitate structural analysis, while optimizing protein expression and solubility. The advantages and disadvantages for various production platforms are discussed, allowing for the best approach to be selected for any particular facility. Potential rescue strategies and quality assurance systems are described.