Proteins: Structure and Function

ISBN-13: 9781461290018 / Angielski / Miękka / 2011 / 834 str.

James J. L'Italien

Opis

This volume surveys the current status of many of the important methods and approaches which are central to the study of protein structure and function. Many of the articles in this volume are written to emphasize the general utility of the method or approach which is at its core, and to provide sufficient literature references to enable the reader to adapt the method or approach to other applications. It is hoped that this volume will provide a source from which newcomers as well as experienced scient- ists may becom& more familiar with recent developments and future trends in some of the important areas of protein research. The articles which comprise this book are selected proceedings from the Symposium of American Protein Chemists, which was held in San Diego, California, September 30 to October 3, 1985. The goal of the organizers of this first symposium was to provide a forum for discussion and inter- action among scientists whose interests span the broad spectrum of protein structure and function research. The concept and timing of the symposium well received as evidenced by the approximately 500 delegates to the was symposium. The inaugural meeting was marked by a strong scientific pro- gram with over 140 papers presented in either a lecture or poster format.

Spis treści

A. Methods of Polypeptide Purification and Characterization.- I. Chromatographic Methods of Polypeptide Purification.- Use of Short HPLC Column For The Concentration, Separation and Recovery of Subnanomole Amounts of Protein and Polypeptides For Microsequence Analysis.- Practical and Theoretical Aspects of Microbore HPLC of Proteins, Peptides and Amino Acid Derivatives.- Microisolation and Sequence Analysis of Human Epidermal Growth Factor.- Complete Amino Acid Sequence Determination of Rat Epidermal Growth Factor:Characterization of a Truncated Form With Full In Vitro Biological Activity.- Use of HPLC Comparative Peptide Mapping in Structure/ Function Studies.- Precolumn Derivatization of Peptides With Phenlisothiocynate (PITC).- High-Performance Liquid Chromatography as a Means of Characterizing Isoforms of Steroid Hormone Receptor Proteins.- Large Scale Purification of a Human Hepatoma Derived Endothelial Cell Growth Factor.- Very Rapid Microanalysis of IgG in Ascites Fluids By HPLC Using a Novel Anion-Exchange Column.- Application of Immunoaffinity Chromatography to the Purification of Polypeptides for Microsequence Analysis.- II. Electrophortic Methods of Polypeptide Purification.- A Novel Approach to Isolation of Proteins for Micro-sequences Analysis:Electroblotting onto Activated Glass.- Blotting of Protein-Detergent Complexes on Glass and Glass Coated with Polybases:A Comparative Study. Acid Hydrolysis and Gas-Phase Sequencing of Gel- Separated Proteins Immobilized on Polybrene Glass Fiber.- Two-Dimensional Gel Electrophoresis of Polypeptides.- III. Chemical Modification of Polypeptides.- Carboxymethylated Hemoglobin as a Structural Analogue For Carbamino Hemoglobin.- Gossypol:Interaction with Ribonuclease A.- Studies on Nonenzymic Glycosylation of Peptides in a Simple Model System.- Reaction of Glyceraldehyde (Aldotriose) with Proteins is a Prototype of Nonenzymic Glycation:Protein Cross-linking as a Consequence of In Vitro Non-Enzymic Glycation.- Reversible Dihydroxypropylation of Amino Groups in Proteins:Application in Primary Structural Studies of Streptococcal M-Proteins.- Enzymatic Carboxylmethylation and Non-Enzymatic Demethylation at L-Isoaspartyl Residues:Possible Applications in Peptide Chemistry.- IV. Amino Acid Analysis of Polypeptides.- A Practical Guide to the General Application of PTC-Amino Acid Analysis.- High Performance Liquid Chromatography of Phenylthiodantoin and Phenylthiocarbamyl Amino Acids.- Amino Acid Analysis Using Pre-Column Derivatization with Phenylisothiocyanate:Matrix Effects and Tryptophan Analysis.- An Improved Reverse Phase HPLC Separation and Post-Column Detection of Amino Acids.- HPLC Assay of Phosphoamino Acids by Fluorescence Detection.- Amino Acid Analysis by Reverse-Phase High Performance Liquid Chromatography:Separation of Phenylthiocarbamyl Amino Acids by Spherisorb Octadecylsilane Columns.- V. Mass Spectrometry of Polypeptides.- Fast Atom Bombardment Mass Spectrometry:Application to Peptide Structural Analysis.- Protein Sequence Determination and FAB Mass Spectrometry.- The Use of HPLC and Mass Spectrometric Techniques to Quantify Endogenous opioid peptides.- Structural Analysis of Proteins of the Nervous System.- Pancreatic Preprosomatostatin Processing:Isolation and Structure of Intermediates and Final Products of Processing.- VI. Microsequence Analysis of Polypeptides by Edman Degraduation.- Progress Toward Polybrene Purification and Utility in Micro-Peptide/Protein Sequencing.- Approaches for Microsequencing:Extraction from SDS gel, Composition, Terminal Analysis.- Manual Gas Phase Microsequencing of Filterchips for Simultaneous Edman Degradation of Multiple Peptide Samples.- Strategies For Sequencing Peptide Mixtures by Selective Blocking.- Amidatioti of Protein Carboxyl Groups.- A Method for Direct Amino Acid Sequence Analysis of the NH2-Terminal Regions of Fibrinogen.- VII. Analysis of PTH Amino Acids.- PTH Amino Acid Analysis.- An On-Line Isocratic HPLC System for the Analysis of PTH-Amino Acids on a Gas-Phase Sequencer.- Routine Analysis of Low-Picomole-Level Phenylthiohy-dantoins by HPLC Using a Diisopropylethylamine Acetate/THF Buffer and Acetonitrile Gradient.- Identification of Side-Chain Protected L -Phenylthiohy- dantoins on Cyano HPLC Columns:An Application to Gas-Phase Microsequencing of Peptides Synthesized on Solid-Phase Supports.- Separation of Amino Acid Phenylthiohydantoin Derivatives by High Pressure Liquid Chromatography.- VIII. Computer Analysis of Protein Sequence Data.- Hydrophobic Moments as Tools for Analysis of Protein Sequences and Structures.- Identification of Protein Surfaces and Interaction Sites by Hydrophilicity Analysis.- The Protein Identification Resource (PIR):An On-Line Computer System for the Characterization of Proteins Based on Comparisons with Previously Characterized Protein Sequences.- Computer Analysis of Protein Sequencing Data.- IX. Miscellaneous Methods of Polypeptide Characterization.- Simultaneous Multiple Peptide Synthesis:The Rapid Preparation of Large Numbers of Peptides.- Mapping Functional Domains of Human Platelet Thrombo-spondin with Electroblotting and High Sensitivity Sequencing.- Immunoprotection - A Novel Approach for Mapping Epitopes on an Antigen.- Micro Method for the Detection of Heparin-Binding Proteins and Peptides.- Rapid and Sensitive Determination of Protein Disulfide Bonds.- Simulation of Protein Hydrodynamic Changes Observed by Urea Gradient Gel Electrophoresis.- B. Analysis of Polypeptide Structure and Function.- X. Site-Directected Mutagenesis.- Site-Specific Mutations in Dihydrofolate Reductase at the Dihydrofolate Binding Site.- Site Saturation Mutagenesis of Active Site Residues of ß-Lactamase.- Enzyme Thermostability and its Enhancement by Genetic Engineering.- Expression of Human C5a in E. coli.- XI. Active Site Studies.- Use of Trinitrobenzene Sulfonate to Determine the pKa Values of Two Active-Site Lysines of Ribulose-bisphosphate Carboxylase/Oxygenase..- Advances in Affinity Labeling of Purine Nucleotide Sites in Dehydrogenases.- Escherichia Coli ADPglucose Synthetase Subtrate Inhibitor Binding Sites Studied by Site(s) Directed Chemical Modification and Mutant Enzyme Characerization.- Active Site and Other Sequence Data from Torpedo Californica Acetylcholinesterase.- Acetylcholine Receptor a-Bungarotoxin Interactions Studied by Chemical Modification, Enzyme Digestion and HPLC.- Imidoesters and the Mechanism of Liver Alcohol Dehydrogenase.- Investigation of Gramicidin Channel Function by Single Amino Acid Replacement Using Non-Genetic Code Amino Acids.- XII. Domain and Topographical Studies.- Identification of Functional Domains of the Inhibitor Protein of cAMP-Dependent Protein Kinase.- Atrial Natriuretic Factor Receptor in Adrenal Plamsa Membrane:Identification by Photo-Affinity Labeling.- Structual Analysis of a 29/38kDa Heparin-Binding Domain of Fibronectin:Evidence That Two Different Subunits of Human Plamsa Fibronectin Arise by Alternative mRNA Splicing.- Structural and Functional Studies on the Purified Insulin Receptor.- Localization and Interaction of Functional Sites on Antithrombin III. Use of an Anti-Hapten Antibody as a Structural Probe.- XIII. Characterization of Proteases.- Properties of the Hydrolase that Catalyzes Removal of the Blocked NH2-Terminal Amino Acid Residues From Polypeptides.- Kinetic Product Analysis of Aspartyl Proteinases Utilizing New Synthetic Substrates and Reversed Phase HPLC.- The Esterase-Like Activity of Covalently Bound C3.- XIV. Identification of Sites of Post-Translateral Modification.- Structural Analysis of Carcinoembryonic Antigen (CEA) and a Related Tumor-Associated Antigen (TEX).- Determination of the Location of NG,NGDimethylarginine in a Glycine-Rich Region of Nucleolar Protein C23.- Structural Characterization of a Murine Lymphocyte Homing Receptor Suggests a Ubiquitinated Branched-Chain Glycoprotein.- XV. New Sequences.- Human C3b/C4b Receptor (CR1):Isolation, Protein Sequence Analysis, and Cloning of a Partial cDNA From Human Tonsil.- HTLV-III/LAV Particle-Associated Proteins:I. Virus Purification, Inactivation and Basic Characterization.- Structure and Function of Fish Antifreeze Polypeptides.- Sequence of a Glycine-Rich Protein From Lizard Claw:Unusual Dilute Acid and Heptafluorobutyric Acid Cleavages.- Primary Structure Studies on Serine Hydroxymethyltransferase.- The Proteins in Avian Eggs:A Test of Protein Separation Methods Based on HPLC.- Amino Terminal Sequence of 7B2:A Novel, Human Pituitary/Brain Polypeptide Present in Gonadotrophs and Released by LHRH.- C. Summary of Symposium Workshop.- The Syposium Test Peptide (STP):Synthesis and Characterization of a Model Peptide.- Workshop in Microanalytical Methods - Experiences With an Unknown Synthetic Peptide.- Contributors.