A General Strategy for Site-specific Incorporation of Unnatural Amino Acids Into Proteins PDF Download
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Author: Spencer Jay Anthony-Cahill
Publisher:
ISBN:
Category :
Languages : en
Pages : 676
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Book Description
Author: Spencer Jay Anthony-Cahill
Publisher:
ISBN:
Category :
Languages : en
Pages : 676
Get Book Here
Book Description
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
A method for the site-specific incorporation of unnatural amino acids into proteins in vivo would significantly facilitate studies of the cellular function of proteins, as well as make possible the biosynthesis of unnatural polymers and proteins with novel structures and activities. Our approach consists of the generation of amber suppressor tRNA/aminoacyl-tRNA synthetase pair that are not catalytically competent with all the endogenous Escherichia coli tRNAs an aminoacyl-tRNA synthetases, followed by directed evolution of such orthogonal aminoacyl-tRNA synthetases to alter their amino acid specificities. A new orthogonal suppressor tRNA/aminoacyl-tRNA synthetase pair in E. coli has been derived from the Saccharomyces cerevisiae tRNA (sub Asp) and aspartyl-tRNA syathetase, and the in vitro and in vivo characteristics of this pair were determined. In order to achieve a high specificity for the amino acid, a direct selection for site-specific incorporation of unnatural amino acids into a reporter epitope displayed on the surface of M13 phage has been developed and characterized. Under simulated selection conditions, phage particles displaying aspartate were enriched over 300-fold from a pool of phage displaying asparagine using monoclonal antibodies raised against the aspartate-containing epitope. The direct phage selection offers very high specificity for the amino acid of interest, Which cannot be achieved by conventional methods.
Author: Kiket Avi Omer Makoubi
Publisher:
ISBN:
Category :
Languages : en
Pages : 506
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Book Description
Author:
Publisher: Academic Press
ISBN: 0080921639
Category : Science
Languages : en
Pages : 334
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Book Description
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, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules Provides a "one-stop shop" for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 55
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Book Description
The goal of this project is to develop methods for the site-specific insertion in vivo of one or more amino acid analogues into proteins in eubacteria and in eukaryotes. The amino acids to be used include those that are photoactivatable, those that are fluorescent, those that carry reactive side chains such as keto groups, heavy atoms such as iodine, those that act as spectroscopic probes, and those that mimic phosphoserine, phosphothreonine, or phosphotyrosine. Besides providing a method for the production of new biomateirals with novel chemical and biological properties, proteins carrying such amino acid analogues will have wide applications in biology. These include studies on the folding, structure, stability and function of proteins, protein-protein interactions, protein localization and protein dynamics in vivo and signal transduction.
Author: James Samuelson
Publisher: Humana Press
ISBN: 9781493962877
Category : Science
Languages : en
Pages : 266
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Book Description
This book provides guidance to those wishing to create enzyme variants. It covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes and the engineering of two difference types of rare-cutting endonucleases.
Author: Nediljko Budisa
Publisher: John Wiley & Sons
ISBN: 3527607099
Category : Science
Languages : en
Pages : 312
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Book Description
The ability to introduce non-canonical amino acids in vivo has greatly expanded the repertoire of accessible proteins for basic research and biotechnological application. Here, the different methods and strategies to incorporate new or modified amino acids are explained in detail, including a lot of practical advice for first-time users of this powerful technique. Novel applications in protein biochemistry, genomics, biotechnology and biomedicine made possible by the expansion of the genetic code are discussed and numerous examples are given. Essential reading for all molecular life scientists who want to stay ahead in their research.
Author: Thomas John Magliery
Publisher:
ISBN:
Category :
Languages : en
Pages : 416
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Book Description
Author: Gustavo Fuertes
Publisher: Frontiers Media SA
ISBN: 2832538029
Category : Science
Languages : en
Pages : 123
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Book Description
The site-specific incorporation of unnatural or non-canonical amino acids (ncAAs) into proteins is a universally important tool for systems bioengineering at the interface of chemistry, biology, and biotechnology. The synergistic use of ncAA and related technologies (e.g. Xeno nucleic acids) should enable: i) New opportunities to manipulate, design and elucidate protein structure, dynamics, and function. ii) A deeper understanding of natural and evolved translational systems and their importance for artificial biology. iii) The synthesis of novel biopolymers, creating a solid basis for synthetic cells, which is also an important technology in the production of new classes of medically relevant protein-based scaffolds. Research on reprogrammed protein translation has now reached an experimental and intellectual maturity: more than 200 ncAA (i.e. more than ten times larger variety than standard amino acids) have been introduced into proteins using different routes: genetic code expansion (GCE), selective pressure incorporation (SPI), chemical mutagenesis, protein semi-synthesis, and peptide synthesis.
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
Our objective is to develop general methods for the site-specific incorporation of amino acid analogues into proteins in bacterial and in eukaryotic cells. The approach consists of the use of an amber suppressor transfer RNA (tRNA) aminoacylated with an amino acid analogue, with the help of a mutant aminoacyl-tRNA synthetase, to insert the amino acid analogue at a specific site in a protein. The site of insertion of the analogue is specified by an appropriately placed amber termination codon within the gene for the protein of interest. This approach has two key requirements: (1) an amber suppressor tRNA, which can not be aminoacylated by any of the endogenous aminoacyltRNA synthetases and (2) an aminoacyi-tRNA synthetase, which aminoacylates the amber suppressor tRNA but no other tRNA in the cell. Therefore, an important first goal is to identify such a 21st aminoacyl-tRNA synthetase tRNA synthetase-amber tRNA pair. This goal has been achieved. Several mutations have been introduced into yeast tyrosyl-tRNA synthetase (TyrRS) for isolating mutants that incorporate iodotyrosine into tRNA instead of tyrosine. Work on an alternative approach applicable to mammalian cells has led to a possible general approach for the introduction of two different amino acid analogues into a protein.
