Metabolons and supramolecular enzyme assemblies /

• Άλλοι συγγραφείς: Schmidt-Dannert, Claudia (Επιμελητής έκδοσης), Quin, Maureen B. (Επιμελητής έκδοσης)
• Μορφή: Ηλ. βιβλίο
• Γλώσσα: English
• Έκδοση: Cambridge, MA : Academic Press, an imprint of Elsevier, 2019.
• Έκδοση: First edition.
• Σειρά: Methods in enzymology ; v. 617.
• Θέματα: Enzymes > Biotechnology.; Enzymology.; Supramolecular chemistry.; Enzymes > metabolism.; Metabolomics > methods.; Metabolic Networks and Pathways.; Metabolome > physiology.; Molecular Biology.; SCIENCE > Chemistry > Industrial & Technical.; TECHNOLOGY & ENGINEERING > Chemical & Biochemical.; Electronic books.

Πίνακας περιεχομένων:

• Front Cover; Metabolons and Supramolecular Enzyme Assemblies; Copyright; Contents; Contributors; Preface; Chapter One: Molecular snapshots of dynamic membrane-bound metabolons; 1. Introduction; 2. SMA copolymer preparation; 2.1. SMA copolymer hydrolysis; 2.1.1. Equipment; 2.1.2. Buffers and reagents; 2.1.3. Procedures; 2.1.4. Notes; 3. Preparation of S. bicolor microsomes; 3.1. Growth of etiolated S. bicolor seedlings; 3.1.1. Equipment; 3.1.2. Buffers and reagents; 3.1.3. Procedures; 3.1.4. Notes; 3.2. Preparation of microsomes; 3.2.1. Equipment; 3.2.2. Buffers and reagents; 3.2.3. Procedures
• 3.2.4. Notes4. Isolation of membrane-bound metabolons from native membranes; 4.1. SMALP preparation from sorghum microsomes; 4.1.1. Equipment; 4.1.2. Buffers and reagents; 4.1.3. Procedures; 4.1.4. Notes; 4.2. Purification of SMALPs containing a target protein; 4.2.1. Equipment; 4.2.2. Buffers and reagents; 4.2.3. Procedures; 4.2.4. Notes; 5. Characterization of SMALPs containing P450 metabolons; 5.1. SMALPs observed with electron microscope; 5.1.1. Equipment; 5.1.2. Buffers and reagents; 5.1.3. Procedures; 5.1.4. Notes; 6. Summary and conclusion; References
• Chapter Two: Tricarboxylic acid metabolon1. Introduction; 1.1. Importance of the TCA metabolon; 1.2. Early evidence of the TCA metabolon and challenges associated with its study; 1.3. Cross-linking mass spectrometric analysis of the TCA metabolon; 2. Preparation of cross-linked native TCA metabolon; 2.1. Isolation of intact mitochondria; 2.2. In situ cross-linking; 2.3. Isolation and characterization of metabolons; 3. Preparation of recombinant TCA metabolon; 3.1. Gene construction for recombinant TCA enzymes; 3.2. Overexpression and purification of recombinant enzymes
• 3.3. In-solution cross-linking and isolation of recombinant metabolons4. Mass spectrometric analysis of the Krebs cycle metabolon; 4.1. In-gel digestion of metabolons; 4.2. Mass spectrometric analysis; 4.3. Mascot database searches; 4.4. Structural reconstitution of the Krebs cycle metabolon; 5. Cross-linking techniques have allowed for the isolation of the intact metabolon; 6. Conclusions; References; Chapter Three: Metabolon formation by chemotaxis; 1. Introduction; 1.1. Metabolon formation; 1.2. Enzyme diffusion; 1.3. Enzyme chemotaxis; 2. Chemotactic assembly of enzyme cascade
• 2.1. Chemotactic assembly in the microfluidic channel2.1.1. Fluorescently labeling enzymes and activity assay; 2.1.2. Microfluidic device fabrication; 2.1.3. Confocal microscope imaging; 2.1.4. Chemotactic migration of hexokinase and aldolase in the microfluidic device; 2.2. Chemotactic colocalization of hexokinase and aldolase; 2.2.1. Colocalization setup; 2.2.2. Analysis of HK and Ald aggregates trajectories; 3. Summary; Acknowledgments; References; Chapter Four: Cloning, expression, and purification of intact polyketide synthase modules; 1. Introduction; 2. Cloning of intact PKS modules