Protein Engineering Protocols

Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general.

Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Divided into two sections, Protein Engineering Protocols reviews rational protein design strategies and directed evolutionary techniques, and their impact on protein engineering.
The first section presents design and computational strategies for protein engineering focusing on designing a protein with desired properties, including examples covering a wide range of engineering techniques, such as protein-protein interactions, DNA binding, antibody mimics, and enzymatic activity. The second section on evolutionary techniques provides step-by-step instructions on library design and statistical assessment of library quality. New methods for DNA shuffling as well as different selection strategies are also presented.
Following the successful Methods in Molecular Biology™ series format, this volume provides a comprehensive guide to the methods used at every stage of the engineering process. By combining a thorough theoretical foundation with detailed protocols, Protein Engineering Protocols will be invaluable to all research workers in the area, from graduate students to senior investigators.