UID:
almafu_9958131653602883
Umfang:
1 online resource (334 p.)
ISBN:
9786612285523
,
9781282285521
,
1282285521
,
9780080921631
,
0080921639
Serie:
Methods in enzymology, v. 462
Inhalt:
By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published
Anmerkung:
Description based upon print version of record.
,
Front Cover; Methods in Enzymology; Copyright Page; Contents; Contributors; Preface; Methods in Enzymology; Chapter 1: Protein Phosphorylation by Semisynthesis: From Paper to Practice; 1. Overview of Protein Phosphorylation; 2.1. Thiophosphate substitution as a phosphomimetic; 2.2. Using amino acid substitution as a phosphomimetic; 4.1. Choosing a protein for semisynthesis; 4.6. C-terminal EPL on an insoluble protein; 4.7. N-terminal semisynthesis; 5.2. Microinjection of phosphonylated enzymes; Acknowledgments; References; Chapter 2: Protein Engineering with the Traceless Staudinger Ligation
,
1. Introduction3.1. Experimental procedure: Synthesis of phosphinothiol I; 5. Preparation of the Phosphinothioester Fragment; 5.1. Experimental procedure: Strategy C1; 5.2. Experimental procedure: Strategy C5; Acknowledgments; References; Chapter 3: Replacement of Y730 and Y731 in the alpha2 Subunit of Escherichia coli Ribonucleotide Reductase with 3-Aminotyrosine using and Evolved Suppressor tRNA/tRNA- Synthetase Pair; 1. Introduction; 3. NH2Y, a Y Analogue for Investigating Enzymatic PCET Reactions; 3.1. Overview: Choice of NH2Y; 3.3. Results; 4. Directed Evolution of NH2Y-RS in E. coli
,
5. Examination of the Fidelity and Specificity of NH2Y Incorporation in a Protein Expressed in E. coli5.1. Overview: The Z-domain as a model; 5.3. Results; 6. Generation of Y730NH2Y-alpha2 and Y731NH2Y-alpha2; 6.3. Successful incorporation of NH2Y into alpha2; 6.4. Protocol for successful expression of NH2Y-alpha2s; 6.5. Results; 6.6. Protocol for purification of NH2Y-alpha2s; 6.8. Protocol for measurement of catalytic activities of NH2Y-alpha2s; 6.10. Results of activity and N3ADP assays; Protocol for determining the EPR and UV-vis properties of NH2Y.-alpha2s; 8. Summary
,
Chapter 4: Semisynthesis of Proteins Using Split Inteins1. Introduction; 2. Protein Splicing in cis and in trans is Performed by Inteins; 3. Design of Split Inteins and Considerations on the Intein Insertion Site; 4. Materials and Methods; 4.1. Solid-phase peptide synthesis of ExN-IntN peptides; 4.2. Construction of IntC-POI plasmids; 4.4. Protein trans-splicing assays; 4.6. Purification and functional analysis of semisynthetic splice products; Acknowledgments; Chapter 5: Expressed Protein Ligation for Metalloprotein Design and Engineering; 1. Introduction
,
2. Methods of Selenocysteine Incorporation3.1. Synthesis of Fmoc-Sec(PMB)-OH; 3.3. Deprotection of selenocysteine-containing 17-mer peptide; 3.4. General procedure for EPL of Azurin(1-111)- Intein-CBD and the C-terminal 17-mer peptide; 3.5. Characterization of selenocysteine azurin; 4. Tuning the Type 1 Copper Reduction Potential Using Isostructural Methionine Analogues; 4.3. General procedure for the peptide cleavage from the resin; 4.4. General procedure for EPL of azurin(1-111)-intein-CBD and the C-terminal peptides containing methionie analogues; References
,
Chapter 6: Using Expressed Protein Ligation to Probe the Substrate Specificity of Lantibiotic Synthetases
,
English
Weitere Ausg.:
ISBN 9780123743107
Weitere Ausg.:
ISBN 0123743109
Sprache:
Englisch
Bookmarklink
