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New approaches for flavin catalysis

Author: Bruce Palfey
Publisher: London : Academic Press, 2019.
Series: Methods in enzymology, v. 620.
Edition/Format:   eBook : Document : EnglishView all editions and formats
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Genre/Form: Electronic books
Material Type: Document, Internet resource
Document Type: Internet Resource, Computer File
All Authors / Contributors: Bruce Palfey
ISBN: 9780128168301 0128168307
OCLC Number: 1100588396
Description: 1 online resource
Contents: Front Cover; New Approaches for Flavin Catalysis; Copyright; Contents; Contributors; Preface; Chapter One: Methods for determining the reduction potentials of flavin enzymes; 1. Introduction; 2. Spectroelectrochemistry method; 3. Xanthine oxidase method; 3.1. Materials; 3.2. Protocol; 3.3. Considerations; 3.4. Data analysis; 4. Effects of ligands; 5. Conclusion; References; Chapter Two: Anaerobic methods for the transient-state study of flavoproteins: The use of specialized glassware to define ... ; 1. Introduction; 1.1. O2 as a substrate or a reactive contaminant 1.2. Categories of flavoprotein reactions2. Controlling the concentration of molecular oxygen; 2.1. The limits of anaerobiosis and approach with glove-boxes; 2.2. The advantages of specialized glassware; 2.3. Instrument design considerations; 2.4. The Schlenk line; 2.5. Anaerobic vessels: Tonometers and cuvettes; 2.6. Exchanging argon for dissolved dioxygen: Making a sample anaerobic; 2.7. Anaerobic preparation of the rapid-mixing instrument; 2.8. Method for mounting tonometers and syringes on the kinetic instrument; 2.9. Gas blending and sparging to control the dioxygen concentration 3. Preparing reduced flavoproteins3.1. Titration with sodium hydrosulfite; 3.2. Double-mixing and reduction using the native reductant; 3.3. Enzymatic reduction using mediators; 3.4. Light mediated reduction; 4. Summary and conclusion; References; Chapter Three: Performing anaerobic stopped-flow spectrophotometry inside of an anaerobic chamber; 1. Introduction; 2. Anaerobic chambers; 2.1. Anaerobic chamber components; 2.2. Maintaining an anaerobic chamber; 2.2.1. Maintaining chamber anaerobiosis; 2.2.2. Maintaining humidity and removing hydrogen sulfide 2.3. Flexible chambers compared to rigid chambers3. Introduction to stopped-flow spectrophotometry; 3.1. Components of a stopped-flow spectrophotometer; 3.2. Anaerobic stopped-flow spectrophotometry; 4. Setting up, preparing, and performing stopped-flow spectrophotometry assays inside of an anaerobic chamber; 4.1. Setting up a stopped-flow spectrophotometer inside of an anaerobic chamber; 4.1.1. Materials; 4.1.2. Procedure; 4.1.3. Notes; 4.2. Setting up a Schlenk line on a cart; 4.2.1. Equipment; 4.2.2. Procedure; 4.2.3. Notes; 4.3. Preparing anaerobic buffers; 4.3.1. Equipment 4.3.2. Buffers and reagents4.3.3. Procedure; 4.3.4. Notes; 4.4. Transferring anaerobic solutions; 4.4.1. Equipment; 4.4.2. Procedure; 4.4.3. Notes; 4.5. Addition of enzymes to maintain anaerobic solutions; 4.5.1. Equipment; 4.5.2. Buffers or reagents; 4.5.3. Procedure; 4.5.4. Notes; 4.6. Determining oxygen concentration in deoxygenated buffers to validate anaerobiosis; 4.6.1. Equipment; 4.6.2. Buffers and reagents; 4.6.3. Procedure; 4.6.4. Notes; 4.7. Preparing anaerobic enzyme solutions; 4.7.1. Equipment; 4.7.2. Buffers and reagents; 4.7.3. Procedure; 4.7.4. Notes
Series Title: Methods in enzymology, v. 620.
Responsibility: Bruce A. Palfey.

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Praise for the Series:"Should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The work most often consulted in the lab." --ENZYMOLOGIA"The Methods in Read more...

 
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