Engineering the Genetic Code is essential reading for all molecular life scientists who want to stay ahead in their research.

Features:

• Includes novel applications in protein biochemistry, genomics, biotechnology and biomedicine made possible by the expansion of the genetic code
• Discusses practical advice for first-time users of this powerful technique
• Explains the different methods and strategies to incorporate new or modified amino acids

Contents

Introduction

• Classical Approaches to Protein Modification
• Peptide Synthesis, Semisynthesis and Chemistry of Total Protein Synthesis
• Chemoselective Ligations Combined with Biochemical Methods
• Methods and Approaches of Classical Protein Engineering
• Genetically Encoded Protein Modifications – Reprogramming Protein Translation
• Basic Definitions and Taxonomy

A Brief History of an Expanded Amino Acid Repertoire

• The ‘‘Golden Years’’ of Molecular Biology and Triplet Code Elucidation
• Early Experiments on the Incorporation of Amino Acid Analogs in Proteins
• Test Tube (Cell-free) Synthesis of Proteins and Early Incorporation Experiments
• Noncanonical Amino Acids as Tools for Studying Cell Metabolism, Physiology, Protein Processing and Turnover
• Problem of Proofs and Formal Criteria for Noncanonical Amino Acid Incorporation
• Recent Renaissance – Genetic Code Engineering

Basic Features of the Cellular Translation Apparatus

• Natural Laws, Genetic Information and the ‘‘Central Dogma’’ of Molecular Biology
• Cellular Investments in Ribosome-mediated Protein Synthesis
• Molecular Architecture of AARS
• Structure and Function of the tRNA Molecule
• Aminoacylation Reaction
• AARS:tRNA Interactions – Identity Sets
• Translational Proofreading
• Ribosomal Decoding – A Brief Overview
• Codon Bias and the Fidelity of Protein Synthesis
• Preprogrammed Context-dependent Recoding: fMet, Sec, Pyl, etc
• Beyond Basic Coding – Posttranslational Modifications

Amino Acids and Codons – Code Organization and Protein Structure

• Basic Features and Adaptive Nature of the Universal Genetic Code
• Metabolism and Intracellular Uptake of Canonical Amino Acids
• Physicochemical Properties of Canonical Amino Acids
• Reasons for the Occurrence of Only 20 Amino Acids in the Genetic Code
• What Properties of Amino Acids are Best Preserved by the Genetic Code
• Evolutionary Legacy: Dual Nature of Conserved Code and Finite Number of Protein Folds
• Natural Variations in Assignment of Codons of the Universal Genetic Code
• Codon Reassignment Concepts Possibly Relevant to Code Engineering

Reprograming the Cellular Translation Machinery

• Enzyme Specificity of Aminoacyl-tRNA Synthetases (AARS) and Code Interpretation
• Reassigning Coding and Noncoding Units
• In Vitro Chemical and Enzymatic tRNA Aminoacylation
• Novel Codon–Anticodon Base Pairs
• Stop Codon Takeover
• In Vivo Nonsense Suppression-based Methods

Implications and Insights: From Reprogrammed Translation and Code Evolution to Artificial Life

• Code Engineering and Synthetic Biology
• Novel Features of Protein Translation that have Emerged from Research in Code Engineering
• The Amino Acid Repertoire and its Evolution
• Artificial Genetic Systems and Code Engineering

Some Practical Potentials of Reprogrammed Cellular Translation

• Practical Choice of Methods and Some Controversies in the Field
• The Plasticity of the Translation Machinery, Amino Acid Generic Types and Protein Structure
• DNA Nucleotide Analogs: From Sequencing to Expanded Code and Therapy
• Noncanonical Amino Acids in Material Science
• Isomorphous Replacement and Atomic Mutations in Structural Biology and Biophysics
• Translationally Active Amino-Trp Analogs: Novel Spectral Windows and Protein Sensors
• Fluorinated Amino Acids in Protein Engineering and Design
• Protein Processing, Bioorthogonality and Protein Surface Diversifications
• Pharmacologically Active Amino Acids

Index

click here to see books • videos • cd-roms of related interest