Flavins and Flavoproteins

Methods and Protocols
1. Auflage 2021
ISBN: 978-1-0716-1288-0
Verlag: Springer US

Buch, Englisch, Band 2280, 298 Seiten, Format (B × H): 178 mm x 254 mm, Gewicht: 590 g

Reihe: Methods in Molecular Biology

This book of protocols is devoted to the yellow coenzymes derived from riboflavin or vitamin B2 and to the hundreds of enzymes whose functionality depends on them, and represents a compendium of techniques for working with flavoproteins or with the wide spectrum of functions that flavoproteins can drive in the cells. Starting with Rf production in microorganisms and the chemical, optical, and redox properties of these fascinating molecules and moving along to the variety and the peculiarity of some single flavoenzymes, the volume explores the complexity of functions and distribution of these molecules in the cell. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 
Authoritative and practical, Flavin and Flavoproteins: Methods and Protocols serves as an ideal guide for protein chemists interested in purifying and characterizing flavoproteins, as well as microbiologists, physiologists, and clinicians, who wish to further study problems connected with flavoproteins.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Barile, Maria

Fachgebiete

• Naturwissenschaften Biowissenschaften Enzymologie
• Naturwissenschaften Chemie Chemie Allgemein

Inhaltsverzeichnis

Selection of Riboflavin Overproducing Strains of Lactic Acid Bacteria and Riboflavin Direct Quantification by Fluorescence.- Recent Advances in Construction of the Efficient Producers of Riboflavin and Flavin Nucleotides (FMN, FAD) in the Yeast Candida Famata.- Overexpression of Riboflavin Excretase Enhances Riboflavin Production in the Yeast Candida Famata.- Functional Study of the Human Riboflavin Transporter 2 Using Proteoliposomes System.- Heterologous Over-Expression of Human FAD Synthase Isoforms 1 and 2.- Purification of Recombinant Human 6His-FAD Synthase (Isoform 2) and Quantitation of FAD / Protein Monomer Ratio by UV/Vis Spectra.- Continuous and Discontinuous Approaches to Study FAD Synthesis and Degradation Catalyzed by Purified Recombinant FAD Synthase or Cellular Fractions.- Redox Titration of Flavoproteins: An Overview.- Anaerobic Stopped-Flow Spectrophotometry with Photodiode Array Detection in the Pre-Steady State: An Application to Elucidate Oxido-Reduction Mechanisms in Flavoproteins.- Atomic Force Microscopy: Single Molecule Imaging and Force Spectroscopy in the Study of Flavoproteins Ligand Binding and Reaction Mechanisms.- Ligand Binding in Allosteric Flavoproteins: Part 1. Quantitative Analysis of the Interaction with NAD of the Apoptosis Inducing Factor (AIF) Harboring FAD in the Reduced State.- Ligand Binding in Allosteric Flavoproteins: Part 2. Quantitative Analysis of the Redox-Dependent Interaction of the Apoptosis-Inducing Factor (AIF) with Its Protein Partner.- Using D- and L-Amino Acid Oxidases to Generate the Imino Acid Substrate to Measure the Activity of the Novel Rid (Enamine/Imine Deaminase) Class of Enzymes.- The In Vitro Production of prFMN for Reconstitution of UbiD Enzymes.- Alcohol Oxidase from the Methylotrophic Yeast Ogataea polymorpha: Isolation, Purification, and Bioanalytical Application.- Flavocytochrome b of the Methylotrophic Yeast Ogataea polymorpha:Construction of Overproducers, Purification, and Bioanalytical Application.- Mammalian Flavoproteome Analysis Using Label-Free Quantitative Mass Spectrometry.- Alteration of Flavin Cofactor Homeostasis in Human Neuromuscular Pathologies.