Methods in Protein Sequence Analysis

Methods in Protein Sequence Analysis contains an intensely prac­ tical account of all the new methodology available to scientists carrying out protein and peptide sequencing studies. Many of the striking advances in fields as diverse as immunology, cell motility, and neurochemistry have in fact been fueled by our ever more powerful ability to determine the sequences and structures of key proteins and peptides. It is our hope that the rich array of tech­ niques and methods for sequencing proteins discussed in this volume-methods that generate much of the information crucial to progress in modern biology-will now become accessible to all who can benefit from them. The papers of the present volume constitute the Proceedings of the IVth International Conference on Methods in Protein Se­ quence Analysis, which was held at Brookhaven National Labo­ ratory, Upton, NY, September 21-25, 1981. It was the most recent in a series of biennial conferences, the previous one having been held in Heidelberg, GFR, in 1979. The series was originated by Richard Laursen, and initially dealt with one aspect of the field, solid-phase sequencing. The scope of the meeting was very broad and among the many aspects of protein sequencing discussed were: instrumentation, strategy, chemicals, mass spectrometry, cleavage of proteins and separation of peptides, and solid, liquid, manual, and even "gas­ phase" sequencing.

Conference Lecture.- An Anecdotal Account of the History of Peptide Stepwise Degradation Procedures.- Overview.- An Evaluation of the Current Status of Protein Sequencing.- Strategy.- Strategic Approaches to Sequence Analysis.- Improvements in Instrumentation and the Chemistry of Isothiocyanate Degradation.- A New Protein Microsequenator Using Gas Phase Edman Reagents.- Toward Longer Degradations on a Sequenator.- The Nature of Non-Specific Peptide Bond Cleavage during the Isothiocyanate Degradation of Proteins.- Microsequencing.- Isolation and Sequence Determination of Polypeptides at the Picomole Level.- Methodologies in the Microsequencing of Proteins and Peptides.- Micro-Sequence Analysis of Ovine Hypothalamic Cortiocotropin Releasing Factor.- Identification of Internal Residues of Lactose Permease of Escherichia coli by Radiolabel Sequencing of Peptide Mixtures.- The Application of Monoclonal Antibodies to the Microsequencing of Proteins.- Solid Phase Sequencing.- Sensitivity Enhancement in Solid Phase Protein Sequence Analysis—An Examination of Methodology.- Progress in Solid-Phase Sequencing: Extended Degradations of Peptides and Proteins.- The Use of DABITC in Automated Solid-Phase Sequencing.- Improved Automatic Conversion for Use with a Liquid-Phase Sequenator.- A Study of the Edman Degradation in the Assessment of the Purity of Synthetic Peptides.- Solid Phase Synthesis of Polypeptides and Oligonucleotides Using a Solid Phase Sequencer.- Manual Sequencing.- Manual Batchwise Sequencing Methods.- Micro Quantitative Edman Manual Sequencing.- Sequencing by Mass Spectrometry.- Protein Sequencing by Mass Spectrometry.- Sequence Analysis of Polypeptides by Direct Chemical Ionization Mass Spectrometry.- A New Computer-Aided Method for Sequencing a Polypeptide from the Masses and Edman-Degradation of Its Constituent Peptide Fragments.- The Complementarity of Mass Spectrometry to Edman Degradation or DNA-based Protein Sequencing.- Cleavage of Proteins.- Enzymic Cleavage of Proteins.- Unusual Enzymatic Cleavage and Preparative Peptide Separation by High Pressure Liquid Chromatography in the Establishment of the Amino Acid Sequence of Tubulin.- Chemical Cleavage of Proteins.- Protein Fragmentation with o-Iodosobenzoic Acid: A Reinvestigation.- Coordination of Protein and DNA Sequencing.- Analysis of the Escherichia coli ATP-Synthase Complex by DNA and Protein Sequencing.- The Primary Structure of Escherichia coli RNA Polymerase. Nucleotide Sequences of the rpo B and rpo C Genes and Amino Acid Sequences of the ? and ?? Subunits.- Correlation between DNA and Protein Sequences in ?-Galactosidase Related Proteins.- The Molecular Consequences of Formaldehyde and Ethyl Methanesulfonate Mutagenesis in Drosophila: Analysis of Mutants in the Alcohol Dehydrogenase Gene.- Use of High Performance Liquid Chromatography (HPLC) For the Separation of Proteins, Peptides, and Amino Acids.- Protein Microsequencing by HPLC Peptide Purification and Solid-Phase Edman Degradation.- Reverse Phase High Performance Liquid Chromatography for Protein Purification: Insulin-like Growth Factors, Ca2+ Binding Proteins and Metallothioneins.- Reversed Phase (RP) HPLC of Proteins and Peptides and Its Impact on Protein Microsequencing.- Preparative Isolation of la Antigen Membrane Protein Component Polypeptides on C18 Reverse Phase HPLC.- Chromatography and Rechromatography in HPLC Separation of Peptides.- Sequence Analysis of Membrane Protein Fragments Isolated by High Pressure Liquid Chromatography.- Preparation of Peptides and Proteins for Sequence Analysis at the Low Nanomole to Subnanomole Level by Reverse-phase High Performance Liquid Chromatography: Results for Cytochromes P450 and Fibronectin.- Analytical and Preparative Applications of High-Performance Liquid Chromatography. Comparisons with Other Methods.- Separation of Hinge Glycopeptides of Human IgD by HPLC.- Use of High Performance Liquid Chromatography in Characterizing Nucleotide Binding Sites and Antigenic Determinants in cAMP-Dependent Protein Kinase.- The Use of Radically-Compressed Reversed Phase HPLC for the Separation of Tryptic Peptides of Apolipoproteins.- Routine Peptide Mapping by High Performance Liquid Chromatography.- Comparative Peptide Mapping by HPLC: Identification of Single Amino Acid Substitutions in Temperature Sensitive Mutants.- A New Low Cost, Fully Automated Amino Acid Analyzer Using a Gradient HPLC.- Identification of Phenylthiohydantoins.- Fused Silica—Capillary—Gas Chromatography. A Novel Procedure for the identification of PTH in the Analysis of Protein Structure.- Communications: Sequence Methods.- Strategies of Sequence Analysis of Regulatory Subunit of Bovine Cardiac Muscle cAMP-Dependent Protein Kinase Type II.- Enhancement of Sensitivity and Reliability of Automated Sequence Analysis.- Manual Sequence Analysis of Retinoid-Binding Proteins.- The Use of HPLC for Structural Studies on Collagen.- NH2-Terminal Sequence of the Clostripain and NBS Peptides of Streptococcal M5 Protein Purified in One Step by HPLC.- High-Speed Liquid Chromatography of PTH-Amino Acids.- Communications: New Sequences.- NADP-Dependent 6-Phosphogluconate Dehydrogenase. Structure of the Enzyme from Sheep Liver.- Sequence Studies on Na, K-ATPase.- N-Terminal Amino Acid Sequences of ?-Gliadins from Bread Wheats.- Recognition of Patterns in Protein Sequences by Metric Analysis.- Amino Acid Sequence of Bovine ?2-Microglobulin.- Sequence Analyses in Characterizations of Functional Properties.- The Primary Structure Determination of Ribosomal Proteins by Combined Amino Acid and Nucleotide Sequencing.- Two Different Spieces of Cytochrome b5 in One Cell.- A Hydrophobic Tryptic Peptide from Bovine Brain.- Comparison of N-Terminal Amino Acid Sequences of ?-Gliadins and a Related Prolamin.- Amino Acid Sequence of a Monomeric Hemoglobin in Heart Muscle of Bullfrog, Rana catesbeiana.- Purification and Micro Sequence Analysis of Active Peptides from Amphibian Skins.- The Amino Acid Sequence of a Novel Decapeptide from the Skin of the Neotropical Frog, Phyllomedusa sauvagei.- Structural and Functional Studies on Nucleolar Protein C23 Using Limited Chemical Cleavage and HPLC.- Function of NAD at the Active Site of Urocanase.- Amino Acid Sequence of the Alpha and Beta Subunits of Ovine Pituitary Follitropin (FSH).- Amino Acid Sequence Diversity Amongst Cytochromes C-556 in the Genus Agrobacterium.