Beschreibung
The purpose of Calpain Methods and Protocols is quite straightf- ward: it is to present the actual experimental methods used in many different laboratories for the study of calpain. It will provide the vital experimental detail, and the discussion of possible pitfalls, for which the standard journals no longer provide space. This will make it as easy as possible for investi- tors interested in calpain to adopt established methods without repeating old mistakes, and to adapt and apply these methods in novel approaches to the many outstanding calpain questions. These questions range from purely biochemical problems of protein structure and enzyme regulation at the molecular level, through large areas of cell biology, to applied and clinical aspects of calpain function in human d- ease. Within this panoply of topics, a wide range of investigators will find many fascinating and as yet unanswered questions about calpain. Calpain Methods and Protocols will provide instant access to many essential te- niques, while saving them the time and effort involved in developing a new method. In addition to questions relating to the normal physiological roles of the calpains, there is considerable evidence that inappropriate calpain activity may have pathological effects in many tissues, for example, following ischemia. This provides a major stimulus for the development of specific calpain inhi- tors for therapeutic purposes, and for the development of methods to evaluate such inhibitors.
Inhaltsverzeichnis
Part I. Purification of Components of the Calpain System. Purification of µ- and m-Calpain and Calpastatin from Animal Tissues, Valery F. Thompson and Darrel E. Goll. A Simple Protocol for Separation and Assay of µ-Calpain, m-Calpain and Calpastatin from Small Tissue Samples, Jan-Olof Karlsson. Purification and Quantification of Calcium-Activated Neutral Proteases I and II and Novel Isoforms from Cultured Osteoblastic Cells by Ion-Exchange Fast Protein Liquid Chromatography, Elsa J. B. Murray, Keyvan Behnam, Mario S. Grisanti, and Samuel S. Murray. Purification of Calpain by Affinity Chromatography on Reactive Red-Agarose or on Casein-Sepharose, Dorothy E. Croall. Affinity Purification of µ-Calpain from Erythrocytes on an Immobilized Peptide from the Plasma Membrane Calcium Pump: Some Studies on Erythrocyte µ-Calpain, Maurizio Molinari and Ernesto Carafoli. Bacterial Expression and Purification of Recombinant m-Calpain, John S. Elce. Purification and Characterization of Crustacean Calpain-Like Proteinases, Donald L. Mykles. Drosophila Calpains: Purification of a Calpain-Like Enzyme from Fruit Flies, and Expression in E. coli, Gaspar Jekely, Marianna Pinter, and Peter Friedrich. Molecular Analysis of p94 and Its Application to Diagnosis of Limb Girdle Muscular Dystrophy Type 2A, Hiroyuki Sorimachi, Yasuko Ono, and Koichi Suzuki. Purification of Recombinant Calpastatin Expressed in Escherichia coli, Masatoshi Maki and Kiyotaka Hitomi. Preparation of Calpastatin Samples for Western Blotting, Masatoshi Maki and Kiyotaka Hitomi. Isolation and Characterization of Calpain Activator Protein from Bovine Brain, Edon Melloni, Mauro Michetti, Franca Salamino, Roberto Minafra, Bianca Sparatore, and Sandro Pontremoli. Part II. Assays of Calpain. Calpain Zymography with Casein or FITC-Casein, J. Simon C. Arthur and Donald L. Mykles. Casein Zymograph Assessment of µ- and m-Calpain Activity After Traumatic Brain Injury in the Rat In Vivo, Xiurong Zhao, Jennifer K. Newcomb, Brian R. Pike, and Ronald L. Hayes. Fluorescence Measurement of Ca2+ Binding to Domain VI of Calpain, J. Simon C. Arthur and John S. Elce. Kinetic Analysis of Human µ-Calpain Autolysis, Peter Tompa and Peter Friedrich. A Sensitive and Continuous Fluorometric Activity Assay Using a Natural Substrate, MAP2, Peter Tompa, Eva Schad, and Peter Friedrich. Measurement of Calpain Activity In Vitro and In Situ Using a Fluorescent Compound and Tau as Substrates, Rodney P. Guttmann and Gail V.W. Johnson. Localization of Calpain by Immunofluorescence in Adherent Cells, Sucheta Kulkarni and Joan E. B. Fox. A Radioimmunologic Technique for Assessing Calpain Activation in Cells, Ronald L. Mellgren. Part III. Specific Tissues. Calpains and Myogenesis, Patrick Cottin, Sylvie Poussard, Elise Dargelos, Denis Balcerzak, Bernadette Aragon, Jean Jacques Brutis, and Andre Ducastaing. Calpastatin (the Endogenous Calpain Inhibitor) and Membrane Protein Degradation in Cell Fusion, Nechama S. Kosower and Sivia Barnoy. The Role of Calpain in Neurofilament Protein Degradation Associated with Spinal Cord Injury, Naren L. Banik and Donald C. Shields. Calpain-Mediated Truncation of Glutamate Ionotropic Receptors: Methods for Studying the Effects of Calpain Activation in Brain Tissue, Xiaoning Bi, Ruifen Bi, and Michel Baudry. Concurrent Assessment of Calpain and Caspase-3 Activity by Means of Western Blots of Protease-Specific Spectrin Breakdown Products, Jennifer K. Newcomb, Brian R. Pike, Xiurong Zhao, and Ronald L. Hayes. Rat Renal Proximal Tubules, Hypoxia, and Calpain, Charles L. Edelstein. Calpain Activity in Rat Renal Proximal Tubules: An In Vitro Assay, Charles L. Edelstein. Calpain Activity in Rat Renal Proximal Tubules: An In Situ Assay, Charles L. Edelstein. Cellular In Vivo Assay of Calpain Activity Using a Fluorescent Substrate: Application to Study of Anoxic Liver Injury, Barry G. Rosser and Gregory J. Gores. Calpain Methods in Hepatic Ischemia-Reperfusion Injury, David Sindram ...
