Publisher's Synopsis
Among all biomolecules in living cells, proteins represent a group with highly divergent biological functions. These include maintaining structural integrity, catalyzing most chemical reactions in vivo, regulating cellular responses via signal transduction, and targeting foreign molecules via the immune system. Because of these diverse functions and associated applications, protein engineering has been a field for intense academic research and biopharmaceutical drug development. Protein engineering, which is the design and construction of novel proteins, usually by manipulation of their genes, is a promising approach which can be used to create enzymes with the desired properties. Proteins are engineered with the goal of better understanding the molecular basis for their functions and also so they will be able to synthesize novel products in non-native environments. Success would greatly expand the possible applications of enzymes in industrial processes. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins (Arnold, 1993). Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first polymerase chain reaction (PCR), where two PCRs take place, and two double-stranded DNA products are obtained. Methods for Protein Engineering presents in-depth discussions of various methods for protein engineering featuring contributions from renowned experts from different counties. It covers significant aspects of methods and applications in the design of novel proteins with different functions. Rational design in other words computational design of proteins requires the amino acid sequence, 3D structure and function knowledge of the protein of interest. This method provides controllable amino acid sequence changes (insertion, deletion or substitution). Controlled changes are important to determine the effect of individual residue changes on the protein structure, folding, stability or function. Knowledge of three-dimensional structure is a key for understanding the biological function. Although understanding of 3D structure of proteins is crucial in terms of their function, only about 1 % of proteins for which the amino acid sequence is known, had their 3D structure determined because of the time consuming nature and difficulty of crystallographic experimental methods. As a result, the gap between the numbers of known sequences and structures continuously grows. In addition to enlarging databases, improvements in sequence comparison, fold recognition and protein modelling algorithms have supported the enhancement of protein structure prediction studies based on computer modelling methods to bridge this gap. Among all biomolecules in living cells, proteins represent a group with highly divergent biological functions. These include maintaining structural integrity, catalyzing most chemical reactions in vivo, regulating cellular responses via signal transduction, and targeting foreign molecules via the immune system. Because of these diverse functions and associated applications, protein engineering has been a field for intense academic research and biopharmaceutical drug development. Protein engineering, which is the design and construction of novel proteins, usually by manipulation of their genes, is a promising approach which can be used to create enzymes with the desired properties. Proteins are engineered with the goal of better understanding the molecular basis for their functions and also so they will be able to synthesize novel products in non-native environments. Success would greatly expand the possible applications of enzymes in industrial processes. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins (Arnold, 1993). Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first polymerase chain reaction (PCR), where two PCRs take place, and two double-stranded DNA products are obtained. Methods for Protein Engineering presents in-depth discussions of various methods for protein engineering featuring contributions from renowned experts from different counties. It covers significant aspects of methods and applications in the design of novel proteins with different functions. Rational design in other words computational design of proteins requires the amino acid sequence, 3D structure and function knowledge of the protein of interest. This method provides controllable amino acid sequence changes (insertion, deletion or substitution). Controlled changes are important to determine the effect of individual residue changes on the protein structure, folding, stability or function. Knowledge of three-dimensional structure is a key for understanding the biological function. Although understanding of 3D structure of proteins is crucial in terms of their function, only about 1 % of proteins for which the amino acid sequence is known, had their 3D structure determined because of the time consuming nature and difficulty of crystallographic experimental methods. As a result, the gap between the numbers of known sequences and structures continuously grows. In addition to enlarging databases, improvements in sequence comparison, fold recognition and protein modelling algorithms have supported the enhancement of protein structure prediction studies based on computer modelling methods to bridge this gap. Among all biomolecules in living cells, proteins represent a group with highly divergent biological functions. These include maintaining structural integrity, catalyzing most chemical reactions in vivo, regulating cellular responses via signal transduction, and targeting foreign molecules via the immune system. Because of these diverse functions and associated applications, protein engineering has been a field for intense academic research and biopharmaceutical drug development. Protein engineering, which is the design and construction of novel proteins, usually by manipulation of their genes, is a promising approach which can be used to create enzymes with the desired properties. Proteins are engineered with the goal of better understanding the molecular basis for their functions and also so they will be able to synthesize novel products in non-native environments. Success would greatly expand the possible applications of enzymes in industrial processes. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins (Arnold, 1993). Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first polymerase chain reaction (PCR), where two PCRs take place, and two double-stranded DNA products are obtained. Methods for Protein Engineering presents in-depth discussions of various methods for protein engineering featuring contributions from renowned experts from different counties. It covers significant aspects of methods and applications in the design of novel proteins with different functions. Rational design in other words computational design of proteins requires the amino acid sequence, 3D structure and function knowledge of the protein of interest. This method provides controllable amino acid sequence changes (insertion, deletion or substitution). Controlled changes are important to determine the effect of individual residue changes on the protein structure, folding, stability or function. Knowledge of three-dimensional structure is a key for understanding the biological function. Although understanding of 3D structure of proteins is crucial in terms of their function, only about 1 % of proteins for which the amino acid sequence is known, had their 3D structure determined because of the time consuming nature and difficulty of crystallographic experimental methods. As a result, the gap between the numbers of known sequences and structures continuously grows. In addition to enlarging databases, improvements in sequence comparison, fold recognition and protein modelling algorithms have supported the enhancement of protein structure prediction studies based on computer modelling methods to bridge this gap.
