Includes cutting-edge methods and protocols
Provides step-by-step readily reproducible results
Contains key notes and implementation advice from the experts
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 Protocols serves as an ideal guide for scientists and contributes to directing the attention of researchers to the many fields of growing scientific interest in non-natural amino acids.
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
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(TM) 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 Protocols serves as an ideal guide for scientists and contributes to directing the attention of researchers to the many fields of growing scientific interest in non-natural amino acids.
Preparation of Unnatural Amino Acids with Ammonia-lyases and 2,3-aminomutases.- Multistep Enzyme Catalyzed Reactions for Unnatural Amino Acids.- Enzymatic Production of Enantiopure Amino Acids from Mono-substituted Hydantoin Substrates.- Preparation of Glutamate Analogues by Enzymatic Transamination.- Carbon-Carbon Bond Forming Enzymes for the Synthesis of Non-natural Amino Acids.- Engineering Cyclic Amidases for Non-natural Amino Acid Synthesis.- NMR Analysis of Unnatural Amino Acids in Natural Antibiotics.- Site-specific Incorporation of Unnatural Amino Acids as Probes for Protein Conformational Changes.- Application of Unnatural Amino Acids to the De novo Design of Selective Antibiotic Peptides.- Use of Unnatural Amino Acids to Probe Structure-activity Relationships and Mode-of-action of Antimicrobial Peptides.- Experimental Methods for Scanning Unnatural Amino Acid Mutagenesis.- Genetic Incorporation of Unnatural Amino Acids into Proteins in Yeast.- Site-specific Incorporation of Unnatural Amino Acids into Proteins in Mammalian Cells.- Incorporation of Unnatural Non-a-amino Acids into the N-terminus of Proteins in a Cell-free Translation System.- Site Specific Modification of Proteins by the Staudinger-phosphite Reaction.- HPLC Methods for Determination of D-aspartate and N-methyl-D-aspartate.- Estimation of Chronological Age from the Racemization Rate of L- and D-aspartic Acid: How to Completely Separate Enantiomers from Dentin.- Enzymatic Detection of D-amino Acids.- An Enzymatic-HPLC Assay to Monitor Endogenous D-serine Release from Neuronal Cultures.- Electrophysiological Analysis of the Modulation of NMDA-receptors Function by d-serine and Glycine in the Central Nervous System.- Biosensors for D-amino Acid Detection.- Analysis of D-beta-aspartyl Isomers at Specific Sites in Proteins.- Nutritional Value of D-amino Acids, D-peptides and Amino Acid Derivatives in Mice.- Preparation and Assay of Recombinant Serine Racemase.- Assay of Amino AcidRacemases.- Assays of D-amino Acid Oxidases.- Enzymes Acting on D-amino Acid Containing Peptides.
Inhaltsverzeichnis
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-ß-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
Klappentext
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.n nAuthoritative and accessible, Unnatural Amino Acids: Methods and Protocols serves as an ideal guide for scientists and contributes 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
