Single Molecule Enzymology: Methods and Protocols

ISBN-13: 9781493961900 / Angielski / Miękka / 2016 / 294 str.

Gregory I. Mashanov; Christopher Batters

Opis

This an ideal guide for the diverse community of molecular biologists, biochemists, and biophysicists studying enzymes in detail. Experts in the field provide procedures which enable the extraction of detailed information about enzyme work cycles.

Spis treści

1. Single Enzyme Studies: A Historical Perspective

            Alex E. Knight

 

2. Functional Surface-Attachment in a Sandwich Geometry of GFP-Labeled Motor Proteins

            Volker Bormuth, Felix Zörgibel, Erik Schäffer, and Jonathon Howard

 

3. Studying Kinesin’s Enzymatic Cycle Using a Single-Motor Confocal Motility Assay, Employing Förster Resonance Energy Transfer

            Zdenek Lansky and Erwin J.G. Peterman

 

4. Fluorescence Imaging with One Nanometer Accuracy: In Vitro and In Vivo Studies of Molecular Motors

            Melinda Tonks Hoffman, Janet Sheung, and Paul R. Selvin

 

5. Snapshots of Kinesin Motors on Microtubule Tracks

            Franck J. Fourniol and Carolyn A. Moores

 

6. Structural and Dynamic Characterization of Biochemical Processes by Atomic Force Microscopy

            Frédéric Eghiaian and Iwan A.T. Schaap

 

7. Using Optical Tweezers to Study Fine Details of Myosin ATPase Mechanochemical Cycle

            Christopher Batters and Claudia Veigel

 

8. Quantum Dot Labeling Strategies to Characterize Single Molecular Motors

            Shane R. Nelson, M. Yusuf Ali, and David M. Warshaw

 

9. Imaging Individual Myosin Molecules Within Living Cells

            Tatiana A. Nenasheva, Gregory I. Mashanov, Michelle Peckham, and Justin E. Molloy

 

10. Single Molecule Measurements Using Microneedles

            Toshio Yanagida, Yoshiharu Ishii, and Akihiko Ishijima

 

11. Fluorescent Nucleoside Triphosphates for Single Molecule Enzymology

            Christopher P. Toseland and Martin R. Webb

 

12. Probing the Mechanics of the Complete DNA Transcription Cycle in Real-Time Using Optical Tweezers

            Christoph G. Baumann and Stephen J. Cross

 

13. A Single-Molecule Approach to Visualize the Unwinding Activity of DNA Helicases

            Natalia Fili, Christopher P. Toseland, Mark S. Dillingham, Martin R. Webb, and Justin E. Molloy

 

14. Real-Time Single-Molecule Observation of Green Fluorescent Protein Synthesis by Immobilized Ribosomes

            Ryo Iizuka, Takashi Funatsu, and Sotaro Uemura

 

15. Single-Molecule Measurements of Topoisomerase Activity with Magnetic Tweezers

            Yeonee Seol and Keir C. Neuman

 

16. Assembly of Recombinant Nucleosomes on Nanofabricated DNA Curtains for Single Molecule Imaging

            Ja Yil Lee and Eric C. Greene

 

17. Simultaneous Observation of Chemomechanical Coupling of a Molecular Motor

            Takayuki Nishizaka*, Yuu Hashimoto, and Tomoko Masaike

 

18. Microsecond Resolution of Single Molecule Rotation Catalyzed by Molecular Motors

            Tassilo Hornung, James Martin, David Spetzler, Robert Ishmukhametov, and Wayne D. Frasch

Blurb

The last fifteen years have witnessed the birth and maturation of many original methods and the development of protocols specific to single molecule measurements and their analysis, including techniques involving optical imaging, electron microscopy, optical and magnetic trapping, and developments in atomic force microscopy.  In Single Molecule Enzymology: Methods and Protocols, experts in the field provide procedures which enable the extraction of detailed information about enzyme work cycles, their static and kinetic properties, and information about their location and activity within cells. The detailed volume offers practical advice on many aspects of single molecule enzymology and includes strategic overviews of interconnected methods involved in sample preparation, single molecule measurements, and data analysis.  Written in the highly successful Methods in Molecular Biology™ series format, chapters contain 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 up-to-date, Single Molecule Enzymology: Methods and Protocols is intended for use within the diverse community of molecular biologists, biochemists, and biophysicists studying enzymes in detail and can be used by researchers planning their first single molecule study or to aid more experienced researchers in further developing their existing studies.