Techniques in Protein Chemistry

Contents

Foreword

Preface

Acknowledgments

Section I Protein Microsequence Analysis

Introduction

1. Strategies for Microcharacterization of Proteins Using Direct

Chemistry on Sequencer Supports

2. Manual Gas-Phase Microsequencing of Peptides and Proteins

3. Microsequencing with PVDF Membranes: Efficient Electroblotting, Direct Protein Adsorption and Sequencer Program Modifications

4. Recovery of Biological Activity, Blotting and Sequence Analysis of Protein Hormones after SDS-PAGE

5. Direct Sequence Analysis of Rat Liver Membrane Glycoproteins Separated by 2D-PAGE

6. The Use of Phenylthiocarbamyl Amino Acid Esters for Sensitivity Enhancement in Edman Degradation

7. Studies on C-Terminal Analysis

8. Studies on the Use of Silyl Compounds for Protein Carboxy-Terminal Sequence Analysis

9. An Improved Injection System for On-Line Liquid Chromatographs: Application to High Sensitivity Phenylthiohydantoin Amino Acid Analysis

10. A Synthetic Peptide for Evaluating Protein Sequencer and Amino Acid Analyzer Performance in Core Facilities: Design and Results

11. Protein Sequencing Sample Advisor: An Expert System to Assess Sample Suitability for Microsequencing

12. Detection of Protein Structural Features with Profile Analysis

Section II Mass Spectrometry and NMR Spectroscopy

Introduction

13. Structural Analysis of Protein Carboxyl Methyltransf erase Utilizing Tandem Mass Spectrometry

14. Sequence Determination of N-Terminally Blocked and Internal Peptides at the Picomole Level by Tandem Mass Spectrometry with Array Detection 135

15. A New Strategy for Mapping the Topography of a Transmembrane Protein Using Mass Spectrometry

16. Laser Photodissociation Fourier Transform Mass Spectrometry: New Methodology for Sequence Analysis of Oligopeptides and Location of Disulfide Bonds

17. Sequence Analysis of Protein C-Terminal Proteolytic Fragments, Protein Isolated from 2D-gels, and Murine Calbindin: New Methodology

18. Location of Disulfide Bonds in Proteins by Partial Acid Hydrolysis and Mass Spectrometry

19. Evaluation of Methods for the Analysis of Disulfide Containing Peptides by Fast Atom Bombardment Mass Spectrometry

20. Examination of the Posttranslational Modifications of Carcinoembryonic Antigen

21. The Assignment of Proton Resonances in 2D NMR Spectra of Proteins

22. Solution Structure of TGFα by 2D ¹NMR and Molecular Modeling

23. Study of a Phosophorylated Protein by Two-Dimensional NMR Spectroscopy

24. Analytical Aspects of 2D 1H NMR: Purity versus Conformational Heterogeneity in Calbindin D9k

Section III Amino Acid Analysis

Introduction

26. Simple, Economical Amino Acid Analysis Based on Pre-column Derivativation with 9-Fluorenylmethyl Chloroformate (FMOC)

27. Successful PTC Amino Acid Analysis at the Picomole Level

28. A Comprehensive Approach to Amino Acid Analysis

29. Automatic Hydrolysis and PTC Amino Acid Analysis: A Progress Report

30. Amino Acid Analysis at the Femtomole Level Using the Dimethylaminoazobenzene Sulfonyl Chloride Precolumn Dérivatization Method: Potential and Limitation

31. Detection of Norleucine Incorporation into Recombinant Proteins Synthesized in E. Colt

32. Occurrence of Methionine Sulfoxide during Production of Recombinant Human Insulin-like Growth Factor I (IGF-I)

Section IV Applications of HPLC

Introduction

33. Techniques in Protein Chemistry Optimization of Monoclonal Antibody Purification

34. New Hydrophilic Polymer for Protein Separations by HPLC

35. Effects of pH and Ion Pairing Agents on the Reverse Phase HPLC of Hydrophilic Peptides: A Case Study of Pentigetide and Its Fragments

36. Chromatographie Analysis of Glycoprotein Derived Carbohydrates: Pellicular Ion Exchange Resins and Pulsed Electrochemical Detection

37. Enzymatic Digestion of Proteins and HPLC Peptide Isolation in the Subnanomole Range

38. The Use of Glass Capillary Tubes as Disposable Microbore Columns for RP-HPLC of Proteins and Peptides

39. Analysis of Peptides at Low Levels Using a Chromatography System Optimized for Narrow-Bore Columns

Section V Protein Chemistry and Analyses

Introduction

40. Rapid Two-Step Purification of Human Placental Estrogen Synthetase by Substrate-Specific Affinity Chromatography

41. Computer Assisted Design of Recognition Peptides

42. Non-Chromatographic Separation of Peptides

43. An Endogenous Bacterial Protease Specifically Cleaves a Fusion Protein Junction Consisting of Consecutive Basic Amino Acids to Generate a Biologically Active Eukaryotic Adhesive Protein, Bindin

44. A Rapid Procedure for the Quantitative Analysis of Monoclonal Antibodies by High Performance Capillary Electrophoresis

45. Factors Affecting Plate Height in High Performance Zonal Capillary Electrophoreses (HPZCE)

46. Analysis of Proteins in Bronchoalveolar Lavage Fluids During Pulmonary Edema Resulting from Nitrogen Dioxide and Cadmium Exposure

47. A New Assay for Enterokinase

48. Meeting the Challenge in Peptide Fragment Purification for Protein Sequencing

49. Fmoc Chemistry: Synthesis, Kinetics, Cleavage and Deprotection of Arginine-Containing Peptides

50. Salt Bridge Conversion to Amide Bonds with C2N2

51. Thermodynamic Bookkeeping

52. Investigation of Helix Dipole and Salt Bridge Stabilization of Synthetic Helical Peptides Using CD and NMR

Section VI Standard Test Peptide

Introduction

53. Design and Synthesis of Symposium Test Peptide-3 (STP-3)

54. Structure Determination of STP-3

55. Structure of Symposium Test Peptide-3

56. Characterization of Symposium Test Peptide

57. Characterization and Synthesis of STP-3

Index