By Hidetoshi Kono, Wei Wang, Jeffery G. Saven (auth.), Katja M. Arndt, Kristian M. Müller (eds.)
Protein engineering is an interesting mix of molecular biology, protein constitution research, computation, and biochemistry, with the objective of constructing invaluable or worthwhile proteins. Divided into sections, Protein Engineering Protocols studies rational protein layout options and directed evolutionary innovations, and their impression on protein engineering.
the 1st part provides layout and computational techniques for protein engineering concentrating on designing a protein with wanted homes, together with examples overlaying quite a lot of engineering thoughts, similar to protein-protein interactions, DNA binding, antibody mimics, and enzymatic task. the second one part on evolutionary thoughts offers step by step directions on library layout and statistical overview of library caliber. New equipment for DNA shuffling in addition to assorted choice ideas also are presented.
Following the profitable tools in Molecular Biology™ sequence structure, this quantity presents a finished consultant to the tools used at each degree of the engineering method. by way of combining an intensive theoretical beginning with exact protocols, Protein Engineering Protocols might be necessary to all examine employees within the zone, from graduate scholars to senior investigators.
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We call this idea the peptide velcro (PV) hypothesis (8), in reference to the design by Kim and co-workers of an obligate heterodimeric coiled coil termed “Peptide Velcro” (9). This pair of peptides is identical except at the e and g positions, where one sequence contains Lys and the other sequence contains Glu (see Note 2). This peptide pair (like other similar pairs) forms a stable heterodimer in vitro. A recent study tested the PV hypothesis by directly comparing rational design and genetic selection strategies (8).
Res. Commun. 39, 13–19. 12. Guan, K. L. and Dixon, J. E. (1991) Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal. Biochem. 192, 262–267. 02_Bacher 7/14/06 34 5:01 PM Page 34 Bacher and Ellington 13. Xu, Z. , Love, M. , Ma, L. , et al. (1989) Tryptophanyl-tRNA synthetase from Bacillus subtilis. Characterization and role of hydrophobicity in substrate recognition. J. Biol. Chem. 264, 4304–4311.
67. Jensen, L. , Andersen, K. , and Kretzschmar, T. (1998) Scoring functions for computational algorithms applicable to the design of spiked oligonucleotides. Nucleic Acids Res. 26, 697–702. 22 Kono et al. 68. Wolf, E. and Kim, P. S. (1999) Combinatorial codons: a computer program to approximate amino acid probabilities with biased nucleotide usage. Protein Sci. 8, 680–688. 69. Wang, W. and Saven, J. G. (2002) Designing gene libraries from protein profiles for combinatorial protein experiments.