Biomolecular applications of single-molecule measurements: kinetics and dynamics of a single-enzyme reaction

2002 ◽  
Author(s):  
Matt Paige ◽  
David P. Fromm ◽  
William E. Moerner
Keyword(s):  
Single Molecule ◽  
Enzyme Reaction ◽  
Kinetics And Dynamics ◽  
Single Enzyme

Single-Molecule Nanocatalysis Reveals Facet-Dependent Catalytic Kinetics and Dynamics of Pallidium Nanoparticles

ACS Catalysis ◽  
2017 ◽  
Vol 7 (4) ◽  
pp. 2967-2972 ◽  
Author(s):  
Tao Chen ◽  
Sheng Chen ◽  
Ping Song ◽  
Yuwei Zhang ◽  
Hongyang Su ◽  
...  
Keyword(s):  
Single Molecule ◽  
Kinetics And Dynamics ◽  
Catalytic Kinetics

scholarly journals Single-molecule chemical reaction reveals molecular reaction kinetics and dynamics

2014 ◽  
Vol 5 (1) ◽  
Author(s):  
Yuwei Zhang ◽  
Ping Song ◽  
Qiang Fu ◽  
Mingbo Ruan ◽  
Weilin Xu
Keyword(s):  
Reaction Kinetics ◽  
Chemical Reaction ◽  
Single Molecule ◽  
Molecular Reaction ◽  
Kinetics And Dynamics

scholarly journals Single-molecule enzymology: What can it tell us?

2003 ◽  
Vol 25 (4) ◽  
pp. 24-27 ◽  
Author(s):  
Clive R. Bagshaw
Keyword(s):  
Enzyme Activity ◽  
Single Molecule ◽  
Enzyme Molecule ◽  
Basic Principles ◽  
Single Enzyme ◽  
Single Molecule Enzymology

In the last decade, several methods have been developed to measure enzyme activity at the level of a single enzyme molecule. This represents a considerable technical feat, but what does it allow us to learn about enzymes? Here, the basic principles are reviewed to show that new forms of heterogeneity in activity may be revealed and evidence gained for rare states that would otherwise be swamped in bulk assays.

Size-dependent catalytic kinetics and dynamics of Pd nanocubes: a single-particle study

2016 ◽  
Vol 18 (32) ◽  
pp. 22494-22502 ◽  
Author(s):  
Tao Chen ◽  
Yuwei Zhang ◽  
Weilin Xu
Keyword(s):  
Single Molecule ◽  
Single Particle ◽  
Catalytic Properties ◽  
Size Dependent ◽  
Single Particle Level ◽  
Kinetics And Dynamics ◽  
Particle Level ◽  
Catalytic Kinetics

Size-dependent catalytic properties at the single-particle level: the size-dependent catalytic kinetics and dynamics of Pd nanocubes were revealed based on the single-molecule nanocatalysis of Pd nanocrystals.

Threshold in a Single Enzyme Reaction System

1978 ◽  
Vol 83 (3) ◽  
pp. 859-862 ◽  
Author(s):  
Ryuichi MATSUNO ◽  
kazuhiro NAKANISHI ◽  
Masatake OHNISHI ◽  
Keitaro HIROMI ◽  
Tadashi KAMIKUBO
Keyword(s):  
Reaction System ◽  
Enzyme Reaction ◽  
Single Enzyme

scholarly journals INSIGHTS IN ENZYME FUNCTIONAL DYNAMICS AND ACTIVITY REGULATION BY SINGLE MOLECULE STUDIES

2013 ◽  
Vol 08 (03n04) ◽  
pp. 137-160 ◽  
Author(s):  
SUNE K. JØRGENSEN ◽  
NIKOS S. HATZAKIS
Keyword(s):  
Single Molecule ◽  
Life Time ◽  
Mathematical Treatment ◽  
Dynamic Information ◽  
Current State ◽  
Functional Dynamics ◽  
Single Molecule Studies ◽  
And Behavior ◽  
Transient Intermediates ◽  
Single Enzyme

The advent of advanced single molecule measurements heralded the arrival of a wealth of dynamic information revolutionizing our understanding of protein dynamics and behavior in ways not deducible by conventional bulk assays. They offered the direct observation and quantification of the abundance and life time of multiple states and transient intermediates in the energy landscape that are typically averaged out in non-synchronized ensemble measurements, thus providing unprecedented insights into complex biological processes. Here we survey the current state of the art in single-molecule fluorescence microscopy methodology for studying the mechanism of enzymatic activity and the insights on protein functional dynamics. We will initially discuss the strategies employed to date, their limitations and possible ways to overcome them, and finally how single enzyme kinetics can advance our understanding on mechanisms underlying function and regulation of proteins. [Formula: see text]Special Issue Comment: This review focuses on functional dynamics of individual enzymes and is related to the review on ion channels by Lu,44 the reviews on mathematical treatment of Flomenbom45 and Sach et al.,46 and review on FRET by Ruedas-Rama et al.41

scholarly journals Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples

2020 ◽  
Vol 6 (11) ◽  
pp. eaay0888 ◽  
Author(s):  
Shingo Sakamoto ◽  
Toru Komatsu ◽  
Rikiya Watanabe ◽  
Yi Zhang ◽  
Taiki Inoue ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Single Molecule ◽  
Biological Samples ◽  
Protein Profiling ◽  
Single Molecule Level ◽  
Multiple Substrates ◽  
Hydrolyzing Enzymes ◽  
Enzyme Molecules ◽  
Related Proteins ◽  
Single Enzyme

We established an ultrasensitive method for identifying multiple enzymes in biological samples by using a multiplexed microdevice-based single-molecule enzymatic assay. We used a paradigm in which we “count” the number of enzyme molecules by profiling their single enzyme activity characteristics toward multiple substrates. In this proof-of-concept study of the single enzyme activity–based protein profiling (SEAP), we were able to detect the activities of various phosphoric ester–hydrolyzing enzymes such as alkaline phosphatases, tyrosine phosphatases, and ectonucleotide pyrophosphatases in blood samples at the single-molecule level and in a subtype-discriminating manner, demonstrating its potential usefulness for the diagnosis of diseases based on ultrasensitive detection of enzymes.

Optical Fiber Microarrays for Chemical and Biological Measurements

2008 ◽  
Vol 1133 ◽  
Author(s):  
David R Walt ◽  
Christopher LaFratta ◽  
Michael Webb ◽  
Zhaohui Li ◽  
Hans-Heiner Gorris ◽  
...  
Keyword(s):  
Single Molecule ◽  
Biological Fluids ◽  
High Sensitivity ◽  
Kinetic Properties ◽  
Fluorescent Product ◽  
Coherent Imaging ◽  
Fiber Arrays ◽  
Stochastic Nature ◽  
Enzyme Molecules ◽  
Single Enzyme

Abstract We have used coherent imaging fiber arrays as a platform for preparing chemical sensors and biosensors. Sensors can be made with spatially-discrete sensing sites for multi-analyte determinations. Micrometer sized sensors have been fabricated by etching the cores of an optical imaging fiber to create microwells and loading them with microspheres. These arrays possess both high sensitivity and reproducibility and can be used for making thousands of measurements simultaneously such as for genetic analysis or for the analysis of complex biological fluids. Both optical and optoelectrochemical arrays have been used for multiplexed sensing. In another scheme, the arrays can be used for single molecule detection. In this format, individual molecules, such as enzymes, can be trapped in the microwells by sealing each microwell with a silicone gasket. The enzyme molecules catalyze the formation of a fluorescent product that can be detected readily. The kinetic properties of hundreds to thousands of single enzyme molecules can be monitored simultaneously using this format. By observing the stochastic nature of the single molecule responses, new mechanistic insights into the fundamental nature of the enzymes can be obtained.

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