Biosynthesis of the Cyclotide MCoTI-II Using an Engineered Intein - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Biosynthesis of the Cyclotide MCoTI-II Using an Engineered Intein write by J. A. Camarero. This book was released on 2006. Biosynthesis of the Cyclotide MCoTI-II Using an Engineered Intein available in PDF, EPUB and Kindle. Cyclotides are an emerging family of naturally occurring circular mini-proteins ({approx}30-40 amino acids) characterized by six conserved Cys residues (forming 3 disulfide bridges) that create a topologically unique structure designated as a cyclic cysteine knot (CCK). The cysteine knot motif, which is embedded within the macrocylic backbone, is described as two disulfide bridges that form a ring that is penetrated by the third disulfide bridge. The cyclic backbone and CCK motif together confer cyclotides with a remarkable stability and resistance to proteolytic, chemical, and thermal degradation. Further, cyclotides are functionally diverse and display a wide range of functions including uterotonic activity, trypsin inhibition, cytotoxicity, neurotensin binding, anti-HIV, antimicrobial, and insecticidal activity. Together, these characteristics make cyclotides attractive candidates for both drug design and agricultural applications, both in their native forms and as molecular scaffolds for the incorporation of novel bioactivities. [1] The ability to manipulate production of cyclotides within biological systems is critical for mutagenesis studies, production of grafted products, and the mass production of cyclotides with novel activities. My adviser's hope is to achieve this capability by employing recombinant DNA expression techniques to generate large combinatorial libraries of cyclotides. The advantage in creating a biosynthetic library (containing {approx}10{sup 6}-10{sup 10} members/library vs. chemically based libraries with typical values ranging from {approx}10{sup 3}-10{sup 5} members/library) is that it can be lead to the in vivo application of biological screening and selection methodologies based on a specific clone's ability to affect certain cellular processes.
Biosynthesis of a Fully Functional Cyclotide Inside Living Bacterial Cells
Biosynthesis of a Fully Functional Cyclotide Inside Living Bacterial Cells - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Biosynthesis of a Fully Functional Cyclotide Inside Living Bacterial Cells write by . This book was released on 2007. Biosynthesis of a Fully Functional Cyclotide Inside Living Bacterial Cells available in PDF, EPUB and Kindle. The cyclotide MCoTI-II is a powerful trypsin inhibitor recently isolated from the seeds of Momordica cochinchinensis, a plant member of cucurbitaceae family. We report for the first time the in vivo biosynthesis of natively-folded MCoTI-II inside live E. coli cells. Our biomimetic approach involves the intracellular backbone cyclization of a linear cyclotide-intein fusion precursor mediated by a modified protein splicing domain. The cyclized peptide then spontaneously folds into its native conformation. The use of genetically engineered E. coli cells containing mutations in the glutathione and thioredoxin reductase genes considerably improves the production of folded MCoTI-II in vivo. Biochemical and structural characterization of the recombinant MCoTI-II confirmed its identity. Biosynthetic access to correctly-folded cyclotides allows the possibility of generating cell-based combinatorial libraries that can be screened inside living cells for their ability to modulate or inhibit cellular processes.
Plant Cyclotides
Plant Cyclotides - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Plant Cyclotides write by . This book was released on 2015-11-24. Plant Cyclotides available in PDF, EPUB and Kindle. Advances in Botanical Research publishes in-depth and up-to-date reviews on a wide range of topics in plant sciences. Currently in its 76th volume, the series features several reviews by recognized experts on all aspects of plant genetics, biochemistry, cell biology, molecular biology, physiology and ecology. Publishes in-depth and up-to-date reviews on a wide range of topics in plant sciences Contains commentary by recognized experts on all aspects of plant genetics, biochemistry, cell biology, molecular biology, physiology, and ecology This volume features reviews of the fast moving field of plant cyclotides
Split Inteins
Split Inteins - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Split Inteins write by Henning D. Mootz. This book was released on 2016-10-07. Split Inteins available in PDF, EPUB and Kindle. This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments.
Cyclic Peptides
Cyclic Peptides - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Cyclic Peptides write by Jesko Koehnke. This book was released on 2017-12-15. Cyclic Peptides available in PDF, EPUB and Kindle. This book provides the reader with a comprehensive view of the state-of-the-art of cyclic peptides, from construction to utility in biology and drug discovery.
