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Part I: General Methods to Study IF Proteins
1. How to Study Intermediate Filaments in Atomic Detail
Anastasia A. Chernyatina, John F. Hess, Dmytro Guzenko, John C. Voss and Sergei V. Strelkov
2. Mechanical Properties of Intermediate Filament Proteins
Elisabeth E. Charrier and Paul A. Janmey
3. Multidimensional Monitoring of Keratin Intermediate Filaments in Cultured Cells and Tissues
Nicole Schwarz, Marcin Moch, Reinhard Windoffer and Rudolf E. Leube
4. Phospho-Specific Antibody Probes of Intermediate Filament (IF) Proteins
Hidemasa Goto, Hiroki Tanaka, Kousuke Kasahara and Masaki Inagaki
5. Assays for Post-Translational Modifications of Intermediate Filament Proteins
Natasha T. Snider and M. Bishr Omary
6. Immunofluorescence and Immunohistochemical Detection of Keratins
Cornelia Stumptner, Margit Gogg-Kamerer, Christian Viertler, Helmut Denk and Kurt Zatloukal
7. High Throughput Screening for Drugs that Modulate Intermediate Filament Proteins
Jingyuan Sun , Vincent E. Groppi , Honglian Gui, Lu Chen, Qing Xie, Li Liu and M. Bishr Omary
8. The Use of Withaferin A to Study Intermediate Filaments
Royce Mohan and Paola BargagnaMohan
9. Assays to Study Consequences of Cytoplasmic Intermediate Filament Mutations: the Case of Epidermal Keratins
Tong San Tan, Yi Zhen Ng, Cedric Badowski, Tram Dang, John E A Common, Lukas Lacina, Ildikó Szeverényi and E Birgitte Lane
10. Using Data Mining and Computational Approaches to Study Intermediate Filament Structure and Function
David A.D. Parry
Part II: Mammalian IF Proteins
11. Isolation and Analysis of Keratins and Keratin Associated Proteins from Hair and Wool
Santanu Deb-Choudhury, Jeffrey E. Plowman and Duane P. Harland
12. Skin Keratins
Fengrong Wang, Abigail Zieman and Pierre A. Coulombe
13. Simple Epithelial Keratins
Pavel Strnad, Nurdan Guldiken, Terhi O. Helenius, Julia O. Misiorek, Joel H. Nyström, Iris A.K. Lähdeniemi, Jonas S.G. Silvander, Deniz Kuscuoglu and Diana M. Toivola
14. Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments
Karen M. Ridge, Dale Shumaker, Amélie Robert, Caroline Hookway, Vladimir I. Gelfand, Paul A. Janmey, Jason Lowery, Ming Guo, David A. Weitz, Edward Kuczmarski, and Robert D. Goldman
15. Strategies to Study Desmin in Cardiac Muscle and Culture Systems
Antigoni Diokmetzidou, Mary Tsikitis, Sofia Nikouli, Ismini Kloukina, Elsa Tsoupri, Stamatis Papathanasiou, Stelios Psarras, Manolis Mavroidis and Yassemi Capetanaki
16. Genetic Manipulation of Neurofilament Protein Phosphorylation
Maria R. Jones, Eric Villalón and Michael L. Garcia
17. α-Internexin and Peripherin: Expression, Assembly, Functions and Roles in Disease
Jian Zhao and Ronald K. H. Liem
18. Studying Nestin and its Inter-Relationship with Cdk5
Julia Lindqvist, Num Wistbacka and John E. Eriksson
19. Synemin: Molecular Features and the Use of Proximity Ligation Assay to Study its Interactions
Madhumita Paul and Omar Skalli
20. Targeting Mitogen-Activated Protein Kinase Signaling in Mouse Models of Cardiomyopathy Caused By Lamin A/C Gene Mutations
Antoine Muchir and Howard J Worman
21. In Vivo, Ex Vivo and in Vitro Approaches to Study Intermediate Filaments in the Eye Lens
M. Jarrin, L. Young, W. Wu, J.M. Girkin and R.A. Quinlan
Part III: Non-Mammalian IF Protein Systems
22. Compartment Specific Phosphorylation of Squid Neurofilaments
Philip Grant and Harish C. Pant
23. Using Xenopus Embryos to Study Transcriptional and Post-Transcriptional Gene Regulatory Mechanisms of Intermediate Filaments
Chen Wang and Ben G. Szaro
24. Intermediate Filaments in Caenorhabditis Elegans
Noam Zuela and Yosef Gruenbaum
25. Mechanical Probing of the Intermediate Filament-Rich C. Elegans Intestine
Oliver Jahnel, Bernd Hoffmann, Rudolf Merkel, Olaf Bossinger and Rudolf E. Leube
26. Using Drosophila for Studies of Intermediate Filaments
Jens Bohnekamp, Diane E. Cryderman, Dylan A. Thiemann, Thomas M. Magin and Lori L. Wallrath
Intermediate Filament Proteins, the latest volume in the Methods in Enzymology series covers all the intermediate filaments in vertebrates and invertebrates, providing a unique understanding of the multiple different tissue-specific intermediate filaments.
This volume also covers the latest methods that are currently being used to study intermediate filament protein function and dynamics. It will be an important companion for any experimentalist interesting in studying this protein family in their cell or organism model system.
- Focuses on intermediate filaments, including the latest information
- Provides an up-to-date understanding of the field
- Contains contributions from the major scientists working and publishing in the field
The audience is the cell biology, physiology and biochemistry communities, with a particular emphasis on those scientists who are interested in the cytoskeleton.
- No. of pages:
- © Academic Press 2016
- 13th January 2016
- Academic Press
- Hardcover ISBN:
- eBook ISBN:
Praise for the Series:
"Should be on the shelves of all libraries in the world as a whole collection." --Chemistry in Industry
"The work most often consulted in the lab." --Enzymologia
"The Methods in Enzymology series represents the gold-standard." --Neuroscience
Bishr Omary is professor and chair of physiology and professor of internal medicine at the University of Michigan Medical School, and an investigator at the Veterans Affairs Ann Arbor Healthcare System. Prior to joining the University of Michigan in 2008, he was at Stanford University for 19 years where he served as Chief of the Division of Gastroenterology and Hepatology, director of its NIH-supported digestive disease center, director of its NIH-supported training grant, and an investigator at the Veterans Affairs Palo Alto Healthcare System. He received his undergraduate degree from George Mason University, his PhD in Chemistry from the University of California (UC) San Diego and his MD from the University of Miami. He completed internal medicine residency training at UC Irvine then gastroenterology fellowship at UC San Diego prior to starting his first faculty position at Stanford University. He has published more than 160 original papers and 30 reviews and commentaries. Most of his publications involved studies pertaining to the function, regulation and disease association of keratin intermediate filament proteins in digestive organs. For example, his laboratory developed genetic disease models and showed that human mutations in keratin polypeptides 8, 18 and 19 (K8/K18/K19) predispose their carriers to acute and chronic liver disease progression. His group identified and characterized several keratin associated proteins, and multiple posttranslational modifications of keratins including phosphorylation, glycosylation, sumoylation, transamidation and acetylation. His group also helped define the importance of keratins and their posttranslational modifications in formation of the hepatocyte inclusions termed Mallory-Denk bodies. More recently, his work extended to drug discovery to identify compounds that could be used as potential therapy for intermediate filament associated diseases. He chaired the Gordon Conference on Intermediate Filaments in 2006 and has served as editor or co-editor of several review series on intermediate filament proteins. He has trained more than 20 postdoctoral fellows, many of whom have secured academic positions.
Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI, USA
Dr. Ronald Liem is a Professor of Pathology and Cell Biology at Columbia University Medical Center. He received his B.A. from Amherst College and his Ph.D. in Biophysical Chemistry at Cornell University. Following post-doctoral work at the University of Pennsylvania and Harvard Medical School, he joined the faculty at New York University School of Medicine. In 1987, he moved to his current position at Columbia University. Dr. Liem first started studying intermediate filaments of the nervous system as a post-doctoral fellow at Harvard. He has published extensively on the neuronal and glial intermediate filaments and identified -internexin as another neuronal intermediate filament, in addition to the neurofilament triplet protein. His laboratory also showed that -internexin was expressed earlier in development than the neurofilament proteins. His interest in neurodegenerative diseases led to a focus on mutations in neurofilaments linked to Charcot-Marie-Tooth disease (CMT), the most commonly inherited neurological disorder. He has developed cell and mouse models for the disease. He has also studied the family of cytoskeletal linker proteins called plakins. Knock-out studies of one of these plakins in mice, called bullous pemphigoid antigen 1 (BPAG1) results in sensory neuron degeneration and disorganized neuronal intermediate filaments. Further studies on the domains and functions of the neuronal isoform of BPAG1, as well as a related plakin MACF1 (Microtubule actin crosslinking factor 1) showed that these two proteins are versatile cytoskeletal linker proteins with isoforms that can interact with various cytoskeletal elements, including intermediate filaments, microtubules and microfilaments. In 1985, Dr. Liem co-organized one of the first international meetings on Intermediate Filaments for the New York Academy of Sciences. Subsequently, in 1990, Dr. Liem was the co-organizer and co-chair of the first Gordon Conference on Intermediate Filaments. He served as Chair in 1992 and this conference is still on-going today. Dr. Liem has been the recipient of a Jacob Javits Award from the NINDS and has published over 150 papers.
Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, USA
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