Covalent Enzyme-Substrate Compounds: Detection and Catalytic Competence Rapid Mix-Quench MALDI-TOF Mass Spectrometry for Analysis of Enzymatic Systems Pre-Steady-State Kinetics of Enzymatic Reactions Studied by Electrospray Mass Spectrometry with On-Line Rapid-Mixing Techniques Trapping of -Glycosidase Intermediates Trapping Covalent Intermediates on ß-Glycosidases 2-Hydroxy-6-keto-nona-2,4-diene 1,9-Dioic Acid 5,6-Hydrolase: Evidence from18O Isotope Exchange for gem-Diol Intermediate Nucleoside-Diphosphate Kinase: Structural and Kinetic Analysis of Reaction Pathway and Phosphohistidine Intermediate Galactose-1-Phosphate Uridylyltransferase: Kinetics of Formation and Reaction of Uridylyl-Enzyme Intermediate in Wild-Type and Specifically Mutated Uridylyltransferases Kinetic Evidence for Covalent Phosphoryl-Enzyme Intermediate in Phosphotransferase Activity of Human Red Cell Pyrimidine Nucleotidases Characterization of (2->6) -Sialyltransferase Reaction Intermediates: Use of Alternative Substrates to Unmask Kinetic Isotope Effects Use of Sodium Borohydride to Detect Acyl-Phosphate Linkages in Enzyme Reactions Evidence for Phosphotransferases Phosphorylated on Aspartate Residue in N-Terminal DXDX(T/V) Motif MurC and MurD Synthetases of Peptidoglycan Biosynthesis: Borohydride Trapping of Acyl-Phosphate Intermediates Transaldolase B: Borohydride Reduction Trapping of Schiff Base Intermediate between Dihydroxyacetone and -Amino Group of Active-Site Lysine Residue Use of NMR and Borohydride Trapping to Provide Evidence for Covalent Enzyme-Substrate Imine Intermediate Detection of Covalent Tetrahedral Adducts by Differential Isotope Shift ¹³C NMR: Acetyl-Enzyme Reaction Intermediate Formed by 3-Hydroxy-3-methylglutaryl-CoA Synthase Detection of Intermediates in Reactions Catalyzed by PLP-Dependent Enzymes: O-Acetylserine Sulfhydrylase and Serine-Glyoxalate Aminotransferase Protein Tyrosine Phosphatase: X-Ray Crystallographic Observation of Cysteinyl-Phosphate Reaction Intermediate GTP:GTP Guanylyltransferase: Trapping Procedures for Detecting and Characterizing Chemical Nature of Enzyme-Nucleotide Phosphoramidate Reaction Intermediate -Glutamyl Thioester Intermediate in Glutaminase Reaction Catalyzed by Escherichia coli Asparagine Synthetase B -Glutamyltranspeptidase and -Glutamyl Peptide Ligases: Fluorophosphonate and Phosphonodifluoromethyl Ketone Analogs as Probes of Tetrahedral Transition State and -Glutamyl-Phosphate Intermediate Stoichiometric Redox Titrations of Complex Metalloenzymes Urate Oxidase: Single-Turnover Stopped-Flow Techniques for Detecting Two Discrete Enzyme-Bound Intermediates Nitric Oxide Synthase: Use of Stopped-Flow Spectroscopy and Rapid-Quench Methods in Single-Turnover Conditions to Examine Formation and Reactions of Heme- O2 Intermediate in Early Catalysis Myeloperoxidase: Kinetic Evidence for Formation of Enzyme-Bound Chlorinating Intermediate Time-Resolved Resonance Raman Spectroscopy of Intermediates in Cytochrome Oxidase Porphobilinogen Deaminase: Accumulation and Detection of Tetrapyrrole Intermediates Using Enzyme Immobilization Adenosylcobalamin-Dependent Glutamate Mutase: Pre-Steady-State Kinetic Methods for Investigating Reaction Mechanism Ribonucleotide Reductase: Kinetic Methods for Demonstrating Radical Transfer Pathway in Protein R2 of Mouse Enzyme in Generation of Tyrosyl Free Radical Galactose Oxidase: Probing Radical Mechanism with Ultrafast Radical Probe Kinetic Characterization of Transient Free Radical Intermediates in Reaction of Lysine 2,3-Aminomutase by EPR Lineshape Analysis Demonstration of Peroxodiferric Intermediate in M-Ferritin Ferroxidase Reaction Using Rapid Freeze-Quench Mössbauer, Resonance Raman, and XAS Spectroscopies A Survey of Covalent, Ionic, and Radical Intermediates in Enzyme-Catalyzed Reactions
The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today—truly an essential publication for researchers in all fields of life sciences.
- Spectroscopic Detection of Reaction Intermediates
- Isotopic and Kenetic Detection of Reaction Intermediates
- Chemical Trapping and Inhibitor Methods for Detecting Reaction Intermediates
Biochemists, biophysicists, pharmacologists, molecular biologists, analytical, organic, and medicinal chemists, and graduate students in these disciplines
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- © Academic Press 2003
- 21st October 2002
- Academic Press
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Daniel Lee Purich has been at the forefront of biochemistry research for more than 25 years. He was awarded the National Institutes of Health Research Career Development Award from 1977–1982, the Plous Teaching Award (the University of California Santa Barbara Campus-Wide Teaching Award) in 1977, has been a member of the National Institutes of Health Biochemistry Study Section from 1982–1985, and a member of the Journal of Biological Chemistry Editorial Board from 1981–1986. He has been a member of the American Society of Biological Chemists, the American Chemical Society, the New York Academy of Sciences, the Biochemical Society, and the American Society for Cell Biology. Dr. Purich is currently a Professor and Chairman of the Department of Biochemistry and Molecular Biology at the Florida College of Medicine. He is the author and editor of numerous scientific publications.
University of Florida, Gainesville, U.S.A.