Divided into convenient sections, this book contributes to directing the reader's attention to the many fields of growing sci...
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Divided into convenient sections, this book contributes to directing the reader's attention to the many fields of growing scientific interest in non-natural amino acids. It includes methods and protocols, and provides step-by-step readily reproducible results. Even though they are present in nature, non-proteinogenic amino acids are usually defined as unnatural or non-natural. Beside their structural diversity, interest in these compounds is due to their occurrence in nature, their biological properties, the analytical aspects, their use as probes, and their incorporation into peptides and proteins, among other reasons. Divided into five convenient sections, Unnatural Amino Acids: Methods and Protocols deals with enzymatic methods used to produce non-natural amino acids, aspects concerning the presence of unnatural amino acids in peptides with antimicrobial properties, genetic incorporation of unnatural amino acids into proteins (yeast and mammalian cells), and detection and quantification of D-amino acids and related enzymes.Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and accessible,Unnatural Amino Acids: Methods and Protocolsserves as an ideal guide for scientists andcontributes to directing the attention of researchers to the many fields of growing scientific interest in non-natural amino acids.
Includes cutting-edge methods and protocols Provides step-by-step readily reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras Inhalt Part I. Synthesis of Unnatural Amino Acids 1. Preparation of Unnatural Amino Acids with Ammonia-lyases and 2,3-aminomutases László Poppe, Csaba Paizs, Klaudia Kovács, Florin-Dan Irimie, and Beáta Vértessy 2. Multistep Enzyme Catalyzed Reactions for Unnatural Amino Acids Paola D'Arrigo and Davide Tessaro 3. Enzymatic Production of Enantiopure Amino Acids from Mono-substituted Hydantoin Substrates Gwynneth F. Matcher, Rosemary A. Dorrington, and Stephanie G. Burton 4. Preparation of Glutamate Analogues by Enzymatic Transamination Thierry Gefflaut, Zeinab Assaf, and Martine Sancelme 5. Carbon-Carbon Bond Forming Enzymes for the Synthesis of Non-natural Amino Acids Pere Clapés, Jesús Joglar, and Mariana Gutiérrez 6. Engineering Cyclic Amidases for Non-natural Amino Acid Synthesis Francisco Javier Las Heras-Vázquez, Josefa María Clemente-Jiménez, Sergio Martínez-Rodríguez, and Felipe Rodríguez-Vico Part II. Application of Unnatural Amino Acids 7. NMR Analysis of Unnatural Amino Acids in Natural Antibiotics Franca Castiglione 8. Site-specific Incorporation of Unnatural Amino Acids as Probes for Protein Conformational Changes Jennifer C. Peeler and Ryan A. Mehl 9. Application of Unnatural Amino Acids to the De novo Design of Selective Antibiotic Peptides Rickey P. Hicks and Amanda L. Russell 10. Use of Unnatural Amino Acids to Probe Structure-activity Relationships and Mode-of-action of Antimicrobial Peptides Alessandro Tossi, Marco Scocchi, Sotir Zahariev, and Renato Gennaro Part III. Use of Unnatural Amino Acids in Protein Synthesis 11. Experimental Methods for Scanning Unnatural Amino Acid Mutagenesis Jia Liu and Ashton T. Cropp 12. Genetic Incorporation of Unnatural Amino Acids into Proteins in Yeast Qian Wang and Lei Wang 13. Site-specific Incorporation of Unnatural Amino Acids into Proteins in Mammalian Cells Nobumasa Hino, Kensaku Sakamoto, and Shigeyuki Yokoyama 14. Incorporation of Unnatural Non-a-amino Acids into the N-terminus of Proteins in a Cell-free Translation System Takahiro Hohsaka 15. Site Specific Modification of Proteins by the Staudinger-phosphite Reaction Paul Majkut, Verena Böhrsch, Remigiusz Serwa, Michael Gerrits, and Christian P. R. Hackenberger Part IV. D-amino Acids: Analysis and Applications 16. HPLC Methods for Determination of D-aspartate and N-methyl-D-aspartate George H. Fisher and Mara Tsesarskaia 17. Estimation of Chronological Age from the Racemization Rate of L- and D-aspartic Acid: How to Completely Separate Enantiomers from Dentin Toshiharu Yamamoto and Susumu Ohtani 18. Enzymatic Detection of D-amino Acids Gianluca Molla, Luciano Piubelli, Federica Volontè, and Mirella S. Pilone 19. An Enzymatic-HPLC Assay to Monitor Endogenous D-serine Release from Neuronal Cultures Inna Radzishevsky and Herman Wolosker 20. Electrophysiological Analysis of the Modulation of NMDA-receptors Function by d-serine and Glycine in the Central Nervous System Fabrice Turpin, Glenn Dallérac, and Jean-Pierre Mothet 21. Biosensors for D-amino Acid Detection Silvia Sacchi, Elena Rosini, Laura Caldinelli, and Loredano Pollegioni 22. Analysis of D--aspartyl Isomers at Specific Sites in Proteins Noriko Fujii and Norihiko Fujii 23. Nutritional Value of D-amino Acids, D-peptides and Amino Acid Derivatives in Mice Mendel Friedman and Carol E. Levin Part V. Enzymes Active on D-amino Acids 24. Preparation and Assay of Recombinant Serine Racemase Florian Baumgart, Clara Aicart-Ramos, and Ignacio Rodríguez-Crespo 25. Assay of Amino Acid Racemases Masumi Katane, Masae Sekine, and Hiroshi Homma 26. Assays of D-amino Acid Oxidases Gabriella Tedeschi, Loredano Pollegioni, and Armando Negri 27. Enzymes Acting on D-amino Acid Containing Peptides Yasuhisa Asano