Protein Targeting Protocols

ISBN-13: 9781617377273 / Angielski / Miękka / 2010 / 501 str.

Mark Van Der Giezen

Opis

In this highly anticipated update of the extremely successful Protein Targeting Protocols, experts from around the world provide the latest protocols on for isolating different organelles and the localization of particular proteins using a variety of methods such as light, confocal, and electron microscopy. Unlike the first edition, however, this volume places emphasis on protein targeting of cellular compartments in both prokaryotic and eukaryotic systems. Authors provide state-of-the-art information on the fast moving fields of import machinery of mitochondria and plastids, as well as a detailed protocol on the movement of protein complexes in bacterial membranes using fluorescent recovery after photobleaching. Bacterial protein targeting protocols using the Sec-system, type-V secretion apparatus, and the Tat-pathway are described, as is a periplasmic targeting protocol. Chapters also include bioinformatics methods to guide readers through the ever-increasing number of in silico tools currently available. Editor Mark van der Giezen has succeeded in including targeting protocols from different systems, including animal, plant, fungal, and protist models, providing insights into the intricate challenges presented by these diverse systems. Authoritative and state-of-the-art, Protein Targeting Protocols, Second Edition, provides useful techniques that will aid scientists working with a variety of organisms.

Spis treści

Contents Preface Contributors Bacterial systems 1 Localisation and mobility of bacterial proteins by confocal microscopy and fluorescence recovery after photobleaching (FRAP) Conrad Mullineaux 2 Membrane protein insertion and secretion in bacteria Jeanine de Keyzer, Martin van der Laan and Arnold J.M. Driessen 3 Identification of autotransporter proteins secreted by type V secretion systems in Gram-negative bacteria Yihfen T. Yen and Christos Stathopoulos 4 Periplasmic targeting of recombinant proteins in Escherichia coli Filipe J. M. Mergulhão and Gabriel A. Monteiro 5 In vitro analysis of the bacterial twin-arginine-dependent protein export Michael Moser, Sascha Panahandeh, Eva Holzapfel and Matthias Müller Eukaryotic systems 6 Nuclear import properties of the sex determining factor SRY Jade K. Forwood, Gurpreet Kaur and David A. Jans 7 The mitochondrial machinery for import of precursor protein, a review. Kip Gabriel and Nikolaus Pfanner 8 A genetic method to identify mitochondrial proteins in living mammalian cells Takeaki Ozawa and Yoshio Umezawa 9 In vitro and in vivo methods to study protein import into plant mitochondria Shashi Bhushan, Pavel F. Pavlov, Charlotta Rudhe and Elzbieta Glaser 10 Protein targeting to mitochondria of Saccharomyces cerevisiae and Neurospora crassa; in vitro and in vivo studies Panagiotis Paptheodorou, Gra_nya Doma_ska and Joachim Rassow 11 A mitosome purification protocol based on Percoll density gradients and its use in validating the mitosomal nature of Entamoeba histolytica mitochondrial Hsp70 Jorge Tovar, Siân S.E. Cox and Mark van der Giezen 12 The chloroplast protein import machinery, a review Friederike Hörmann, Jürgen Soll and Bettina Bölter 13 Import of plastid precursor proteins into pea chloroplasts Jürgen Soll and Bettina Bölter 14Protein targeting in 'secondary' or 'complex' chloroplasts Balbir K. Chaal and Beverley R. Green 15 Protein trafficking to the complex chloroplasts of Euglena Rostislav Vacula, Silvia Sláviková and Steven D. Schwartzbach 16 Fluorescent protein fusions for protein localization in plants John Runions, Chris Hawes and Smita Kurup 17 Genetic transformation; a tool to study protein targeting in diatoms Peter Kroth 18 Identification of proteins targeted into the endoplasmic reticulum by cDNA library screening Takeaki Ozawa and Yoshio Umezawa 19 Trafficking through the early secretory pathway of mammalian cells Theresa H. Ward 20 Studying protein export from the endoplasmic reticulum in plants Sally L. Hanton, Loren A. Matheson and Federica Brandizzi 21 Imaging the Golgi apparatus in living mitotic cells Nihal Altan-Bonnet 22 Evaluating yeast biosynthetic vacuolar transport Brian A. Davies, Darren S. carney and Bruce F. Horazdovsky 23 Targeting to lysosomes in mammalian cells: the biosynthetic and endocytic pathways Ann H. Erickson and Jeffrey P. Bocock 24 Monitoring autophagy in yeast: the Pho8_60 assay Daniel J. Klionsky 25 Protein targeting to yeast peroxisomes Ida van der Klei and Marten Veenhuis 26 Expression, localization and topology of fluorescently tagged plasma membrane-targeted transmembrane proteins Jeanne Shepshelovich and Koty Hirschberg General methods 27 Intracellular protein localization by immunoelectron microscopy Tetsuaki Osafune and Steven D. Schwartzbach 28 Separation of proteins by blue native electrophoresis (BN-PAGE) Olga Randelj, Joachim Rassow and Christian Motz 29 Computational prediction of subcellular localization Kenta Nakai and Paul Horton 30 Phylogenetic analysis to uncover organellar origins of nuclear-encoded genes Bernardo Foth