RNA and Protein Synthesis

RNA and Protein Synthesis

1st Edition - January 28, 1981

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  • Editor: Kivie Moldave
  • eBook ISBN: 9780323142342

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Description

RNA and Protein Synthesis is a compendium of articles dealing with the assay, characterization, isolation, or purification of various organelles, enzymes, nucleic acids, translational factors, and other components or reactions involved in protein synthesis. One paper describes the preparatory scale methods for the reversed-phase chromatography systems for transfer ribonucleic acids. Another paper discusses the determination of adenosine- and aminoacyl adenosine-terminated sRNA chains by ion-exclusion chromatography. One paper notes that the problems involved in preparing acetylaminoacyl-tRNA are similar to those found in peptidyl-tRNA synthesis, in particular, to the lability of the ester bond between the amino acid and the tRNA. Another paper explains a new method that will attach fluorescent dyes to cytidine residues in tRNA; it also notes the possible use of N-hydroxysuccinimide esters of dansylglycine and N-methylanthranilic acid in the described method. One paper explains the use of membrane filtration in the determination of apparent association constants for ribosomal protein-RNS complex formation. This collection is valuable to bio-chemists, cellular biologists, micro-biologists, developmental biologists, and investigators working with enzymes.

Table of Contents


  • List of Contributors

    Foreword

    Preface

    Section I. Transfer RNA

    1. Reversed-Phase Chromatography Systems for Transfer Ribonucleic Acids-Preparatory-Scale Methods

    2. Miniature Reversed-Phase Chromatography Systems for the Rapid Resolution of Transfer RNA's and Ribosomal RNA's

    3. Transfer RNA Isoacceptors in Cultured Chinese Hamster Ovary Cells

    4. The Use of BD-Cellulose in Separating Transfer RNA's

    5. Fractionation of Rabbit Liver Methionyl-tRNA Species

    6. RNase Hydrolysis of Aminoacyl-sRNA to Aminoacyl Adenosine

    7. Determination of Adenosine- and Aminoacyl Adenosine-Terminated sRNA Chains by Ion-Exclusion Chromatography

    8. Enzymic Modification of the C-C-A Terminus of tRNA

    9. Methods for Determining the Extent of tRNA Aminoacylation in Vivo in Cultured Mammalian Cells

    10. The Chemical Preparation of Acetylaminoacyl-tRNA

    11. The Synthesis of Oligopeptidyl-tRNA

    12. Preparation of Aminoacyl-Oligonucleotides and Their Binding to Ribosomes

    13. Chemical Modifications of Amino Acids Esterified to tRNA (Other Than Acylations)

    14. Use of in Vitro 32P Labeling in the Sequence Analysis of Nonradioactive tRNAs

    15. A New Method for Attachment of Fluorescent Probes to tRNA

    16. Attachment of Cross-Linking Reagents to tRNA for Protein Affinity Labeling Studies

    17. Enzymatic Synthesis of Trinucleoside Diphosphates of Known Sequence

    18. Use of Purified Isoacceptor tRNAs for the Study of Codon-Anticodon Recognition in Vitro with Sequenced Natural Messenger RNA

    19. Rabbit Liver tRNA Nucleotidyltransferase

    20. Bacterial tRNA Methyltransferases

    21. Assay for AA-tRNA Recognition by the EFTu-GTP Complex of Escherichia coli

    22. Mapping the Structure of Specific Protein-Transfer RNA Complexes by a Tritium Labeling Method

    Section II. Aminoacyl-tRNA Synthetases

    23. Aminoacyl-tRNA Synthetase Complex from Rat Liver

    24. Kinetic Techniques for the Investigation of Amino Acid:tRNA Ligases (Aminoacyl-tRNA Synthetases, Amino Acid Activating Enzymes)

    25. Kinetics of Aminoacyl-tRNA Synthetases Catalyzed ATP-PP Exchange

    26. Applications of Kinetic Methods to AminoacyltRNA Synthetases

    Section III. Polypeptide Chain Initiation

    27. Purification of Protein Synthesis Initiation Factors IF-1, IF-2, and IF-3 from Escherichia coli

    28. Protein Synthesis Initiation Factors from Rabbit Reticulocytes: Purification, Characterization, and Radiochemical Labeling

    29. Initiation Factors for Protein Synthesis from Wheat Germ

    30. Assay of the Binding of the Ternary Complex Met-tRNAf · eIF-2 · GTP to the 40 S Ribosomal Subunit by Sucrose Gradient and CsCl Gradient Analysis

    31. Prokaryotic Ribosome Binding Sites

    32. Characterization of Translational Initiation Regions from Eukaryotic Messenger RNAs

    33. Determination of the Number of Ribosomal Binding Sites on the RNAs of Eukaryotic Viruses

    34. Methods for Analyzing Messenger Discrimination in Eukaryotic Initiation Factors

    35. Isolation of Protein Kinases from Reticulocytes and Phosphorylation of Initiation Factors

    Section IV. Polypeptide Chain Elongation

    36. Isolation of the Protein Synthesis Elongation Factors EF-Τu, EF-Ts, and EF-G from Escherichia coli

    37. Preparation of Homogeneous Escherichia coli Translocation Factor G

    38. The Binding of Escherichia coli Elongation Factor G to the Ribosome

    39. GTP-Dependent Binding of Aminoacyl-tRNA to Escherichia coli Ribosomes

    40. Polypeptide Chain Elongation Factors from Pig Liver

    41. Separation of Cytoplasmic and Mitochondrial Elongation Factors from Yeast

    42. Soluble Protein Factors and Ribosomal Subunits from Yeast. Interaction with Aminoacyl-tRNA

    43. Separation and Characterization of Yeast Elongation Factors

    Section V. Polypeptide Chain Termination

    44. Release Factors: In Vitro Assay and Purification

    45. Mammalian Release Factor: In Vitro Assay and Purification

    Section VI. Ribosomes

    46. Sedimentation Velocity Analysis in Accelerating Gradients

    47. Preparation of Ribosomal Subunits by Large-Scale Zonal Centrifugation

    48. A Gel Electrophoretic Separation of Bacterial Ribosomal Subunits with and without Protein SI

    49. Reconstitution of Ribosomes from Subribosomal Components

    50. Total Reconstitution of 50 S Subunits from Escherichia coli Ribosomes

    51. Purification of Ribosomal Proteins from Escherichia coli

    52. Purification of Ribosomal Proteins from Escherichia coli under Nondenaturing Conditions

    53. Two-Dimensional Polyacrylamide Gel Electrophoresis for Separation of Ribosomal Proteins

    54. A Modified Two-Dimensional Gel System for the Separation and Radioautography of Microgram Amounts of Ribosomal Proteins

    55. Analytical Methods for Ribosomal Proteins of Rat Liver 40 S and 60 S Subunits by "Three-Dimensional" Acrylamide Gel Electrophoresis

    56. Cross-Linking of Ribosomes Using 2-Iminothiolane (Methyl 4-Mercaptobutyrimidate) and Identification of Cross-Linked Proteins by Diagonal Polyacrylamide/Sodium Dodecyl Sulfate Gel Electrophoresis

    57. Photoaffinity Labeling of Ribosomes

    58. Radioactive Chemical Labeling of Ribosomal Proteins and Translational Factors in Vitro

    59. Peptidyl-Puromycin Synthesis on Polyribosomes from Escherichia coli

    60. Separation of Large Quantities of Eukaryotic Ribosomal Subunits by Zonal Ultracentrifugation

    61. Two-Dimensional Polyacrylamide Gel Electrophoresis of Eukaryotic Ribosomal Proteins

    62. Isotopic Labeling and Analysis of Phosphoproteins from Mammalian Ribosomes

    63. Phosphorylation of Ribosomal Proteins: Preparation of Rat Liver Ribosomal Protein Kinases. Assay of the Phosphorylation of Ribosomal Subunit Proteins. Assay of the Function of Phosphorylated Ribosomes

    64. Protein-RNA Interactions in the Bacterial Ribosome

    65. The Use of Membrane Filtration to Determine Apparent Association Constants for Ribosomal Protein-RNA Complex Formation

    66. Preparation and Analysis of Mitochondrial Ribosomes

    Index

Product details

  • No. of pages: 830
  • Language: English
  • Copyright: © Academic Press 1981
  • Published: January 28, 1981
  • Imprint: Academic Press
  • eBook ISBN: 9780323142342

About the Editor

Kivie Moldave

Affiliations and Expertise

Department of Molecular Biology and Biochemistry, University of California, Irvine, California, U.S.A.

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