This volume attempts to describe the purification and assay of the diverse group of proteins involved in otoplasmic organization. Some of the more promising attempts to apply techniques of molecular biology, genetics, and molecular cytochemistry to these proteins are discussed.

Table of Contents:
Preparation of Myosin- and Actomyosin-Related Proteins:H. Swanljung-Collins and J.H. Collins, Rapid, High-Yield Purification of Intestinal Brush Border Myosin I.T.J. Lynch, H. Brzeska, I.C. Baines, and E.D. Korn, Purification of Myosin I and Myosin I Heavy Chain Kinase from Acanthamoeba castellanii.Q.G. Medley, S.-F. Lee, and G.P. C~acot~aae, Purification and Characterization of Myosin II Heavy Chain Kinase A from Dictyostelium.M.A. Conti and R.S. Adelstein, Purification and Properties of Myosin Light Chain Kinases.L.J. Wuestehube, D.W. Speicher, A. Shariff, and E.J. Luna, F-Actin Affinity Chromatography of Detergent-Solubilized Plasma Membranes: Purification and Initial Characterization of Ponticulin from ~Dictyostelium discoideum.J.R. Glenney, Jr. , Purification of Calpactins I and II and Isolation of N-Terminal Tail of Calpactin I.A.R. Bresnick and J. Condeelis, Isolation of Actin-Binding Proteins from Dictyostelium discoideum.M. Fechheimer and R. Furukawa, Preparation of 30,000-Da Actin Cross-Linking Protein from Dictyostelium discoideum.P.A. Janmey, Polyproline Affinity Method for Purification of Platelet Profilin and Modification with Pyrene-Maleimide.M. Rozycki, C.E. Schutt, and U. Lindberg, Affinity Chromatography-Based Purification of Profilin: Actin.M.R. Bubb and E.D. Korn, Purification of Actobindin from Acanthamoeba castellanii.J.R. Bamburg, L.S. Minamide, T.E. Morgan, S.M. Hayden, K.A. Giuliano, and A. Koffer, Purification and Characterization of Low-Molecular-Weight Actin-Depolymerizing Proteins from Brain and Cultured Cells.J.F. Casella and J.A. Cooper, Purification of Cap Z from Chicken Skeletal Muscle.Preparation of Microtubules, Microtubule-Associated Proteins, and Microtubule Motors:M.C. Wagner, K.K. Pfister, S.T. Brady, and G.S. Bloom, Purification of Kinesin from Bovine Brain and Assay of Microtubule-Stimulated ATPase Activity.D.D. Hackney, Isolation of Kinesin Using Initial Batch Ion Exchange.B.M. Paschal, H.S. Shpetner, and R.B. Vallee, Purification of Brain Cytoplasmic Dynein and Characterization of Its in Vitro Properties.H.S. Shpetner and R.B. Vallee, Purification and Characterization of Dynamin.A.G. Moss, J.-L. Gatti, S.M. King, and G.B. ~Witman, Purification and Characterization of Salmo gairdneri Outer Arm Dynein.A.T. Hastie, Purification and Characterization of Dynein from Pig Tracheal Cilia.D.B. Murphy, Purification of Tubulin and Tau from Chicken Erythrocytes: Tubulin Isotypes and Mechanisms of Microtubule Assembly.C.A. Collins, Reversible Assembly Purification of Taxol-Treated Microtubules.S.J. Chapin and J.C. Bulinski, Preparation and Functional Assay of Pure Populations of Tyrosinated and Detyrosinated Tubulin.M. LeDizet and G. Piperno, Detection of Acetylated ~ga-Tubulin by Specific Antibodies.E. Aamodt, Isolation of Microtubules, Adligin, and Other Microtubule-Associated Proteins from Caenorhabditis elegans.D. Robinson, P. Beattie, T. Sherwin, and K. Gull, ~Microtubules, Tubulin, and Microtubule-Associated Proteins of Trypanosomes.Molecular and Genetic Approaches:K.G. Miller, C.M. Field, B.M. Alberts, and D.R. Kellogg, Use of Actin Filament and Microtubule Affinity Chromatography to Identify Proteins That Bind to the Cytoskeleton:T.T. Egelhoff, M.A. Titus, D.J. Manstein, K.M. Ruppel, and J.A. Spudich, Molecular Genetic Tools for Study of the Cytoskeleton in Dictyostelium.E. Wallraff and G. Gerisch, Screening for Dictyostelium Mutants Defective in Cytoskeletal Proteins by Colony Immunoblotting.R. Kamiya, Selection of Chlamydomonas Dynein Mutants.T.C. Huffaker and A.P. Bretscher, Cloning and Analyzing Genes Encoding Cytoskeletal Proteins in Yeast Saccharomyces cerevisiae.E. McNally, R. Sohn, S. Frankel, and L. Leinwand, Expression of Myosin and Actin in Escherichia coli.G.-Q. Xu and S.E. Hitchcock-DeGregori, Synthesis of cDNAs from Synthetic Oligonucleotides Using Troponin C as an Example.Mechanochemical Properties of Motor Proteins:S.J. Kron, Y.Y. Toyoshima, T.Q.P. Uyeda, and J.A. Spudich, Assays for Actin Sliding Movement over Myosin-Coated Surfaces.S. Ishijima and G.B. Witman, Demembranation and Reactivation of Mammalian Spermatozoa from Golden Hamster and Ram.I.R. Gibbons and G. Mocz, Photocatalytic Cleavage of Proteins with Vanadate and Other Transition Metal Complexes.C.R. Cremo, J.C. Grammer, and R.G. Yount, Vanadate-Mediated Photocleavage of Myosin.S.M. King, H. Kim, and B.E. Haley, Strategies and Reagents for Photoaffinity Labeling of ~Mechanochemical Proteins.Chemical Modifications and Assembly Assays:S.R. Heidemann, Microtubule Polarity Determination Based on Formation of Protofilament Hooks.A. Hyman, D. Drechsel, D. Kellogg, S. Salser, K. Sawin, P. Steffen, L. Wordeman, and T. Mitchison, Preparation of Modified Tubulins.J. Condeelis and A.L. Hall, Measurement of Actin Polymerization and Cross-Linking in Agonist-Stimulated Cells.Y.-L. Wang, Preparation and Characterization of Tetramethylrhodamine-Labeled Myosin.K.L. Vikstrom, R.K. Miller, and R.D. Goldman, Analyzing Dynamic Properties of Intermediate Filaments.Each chapter includes references.Author Index.Subject Index.