Split Inteins Methods and Protocols
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 e
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Henning D. Mootz Editor
Split Inteins Methods and Protocols
METHODS
IN
MOLECULAR BIOLOGY
Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
Split Inteins Methods and Protocols
Edited by
Henning D. Mootz Institute of Biochemistry, Department of Chemistry and Pharmacy, University of Muenster, Münster, Germany
Editor Henning D. Mootz Institute of Biochemistry Department of Chemistry and Pharmacy University of Muenster Münster, Germany
ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-6449-9 ISBN 978-1-4939-6451-2 (eBook) DOI 10.1007/978-1-4939-6451-2 Library of Congress Control Number: 2016946364 © Springer Science+Business Media New York 2017 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. Printed on acid-free paper This Humana Press imprint is published by Springer Nature The registered company is Springer Science+Business Media LLC New York
Preface Protein splicing performed by inteins is an exceptional rearrangement of the polypeptide backbone of a precursor protein, in which the intein domain removes itself under concomitant linkage of the flanking extein sequences. The reaction follows a remarkable reaction mechanism consisting of typically three covalent intermediates with reactive thioester or ester linkages. First discovered in 1990 [1, 2], countless protein-based applications of protein splicing have been reported until today in the fields of protein biochemistry, biotechnology, chemical biology, and cell biology. These applications are based on both the changes in structure of the backbone connectivity during splicing and the interception of the reactive intermediates in targeted cleavage reactions. Split inteins represent a subclass of inteins with the intein domain being split into two parts, the N-terminal and C-terminal fragments (IntN and IntC). The fact that the two flankin
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