Single-Molecule Mechanistic Study of Enzyme Hysteresis

Yu Jiang; Xiang Li; Barrett R. Morrow; Arti Pothukuchy; Jimmy Gollihar; Richard Novak; Charles B. Reilly; Andrew D. Ellington; David R. Walt

doi:10.1021/acscentsci.9b00718

Single-Molecule Mechanistic Study of Enzyme Hysteresis

Yu Jiang, Xiang Li, Barrett R. Morrow, Arti Pothukuchy, Jimmy Gollihar, Richard Novak, Charles B. Reilly, Andrew D. Ellington, David R. Walt

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

Hysteresis is an important feature of enzyme-catalyzed reactions, as it reflects the influence of enzyme regulation in the presence of ligands such as substrates or allosteric molecules. In typical kinetic studies of enzyme activity, hysteretic behavior is observed as a "lag" or "burst" in the time course of the catalyzed reaction. These lags and bursts are due to the relatively slow transition from one state to another state of the enzyme molecule, with different states having different kinetic properties. However, it is difficult to understand the underlying mechanism of hysteresis by observing bulk reactions because the different enzyme molecules in the population behave stochastically. In this work, we studied the hysteretic behavior of mutant β-glucuronidase (GUS) using a high-throughput single-molecule array platform and investigated the effect of thermal treatment on the hysteresis.

Original language	English (US)
Pages (from-to)	1691-1698
Number of pages	8
Journal	ACS Central Science
Volume	5
Issue number	10
DOIs	https://doi.org/10.1021/acscentsci.9b00718
State	Published - Oct 23 2019

ASJC Scopus subject areas

Chemistry(all)
Chemical Engineering(all)

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10.1021/acscentsci.9b00718

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title = "Single-Molecule Mechanistic Study of Enzyme Hysteresis",

abstract = "Hysteresis is an important feature of enzyme-catalyzed reactions, as it reflects the influence of enzyme regulation in the presence of ligands such as substrates or allosteric molecules. In typical kinetic studies of enzyme activity, hysteretic behavior is observed as a {"}lag{"} or {"}burst{"} in the time course of the catalyzed reaction. These lags and bursts are due to the relatively slow transition from one state to another state of the enzyme molecule, with different states having different kinetic properties. However, it is difficult to understand the underlying mechanism of hysteresis by observing bulk reactions because the different enzyme molecules in the population behave stochastically. In this work, we studied the hysteretic behavior of mutant β-glucuronidase (GUS) using a high-throughput single-molecule array platform and investigated the effect of thermal treatment on the hysteresis.",

author = "Yu Jiang and Xiang Li and Morrow, {Barrett R.} and Arti Pothukuchy and Jimmy Gollihar and Richard Novak and Reilly, {Charles B.} and Ellington, {Andrew D.} and Walt, {David R.}",

note = "Funding Information: This work is funded by Brigham and Women{\textquoteright}s Hospital and the Wyss Institute. Publisher Copyright: Copyright {\textcopyright} 2019 American Chemical Society.",

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doi = "10.1021/acscentsci.9b00718",

language = "English (US)",

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T1 - Single-Molecule Mechanistic Study of Enzyme Hysteresis

AU - Jiang, Yu

AU - Li, Xiang

AU - Morrow, Barrett R.

AU - Pothukuchy, Arti

AU - Gollihar, Jimmy

AU - Novak, Richard

AU - Reilly, Charles B.

AU - Ellington, Andrew D.

AU - Walt, David R.

PY - 2019/10/23

Y1 - 2019/10/23

N2 - Hysteresis is an important feature of enzyme-catalyzed reactions, as it reflects the influence of enzyme regulation in the presence of ligands such as substrates or allosteric molecules. In typical kinetic studies of enzyme activity, hysteretic behavior is observed as a "lag" or "burst" in the time course of the catalyzed reaction. These lags and bursts are due to the relatively slow transition from one state to another state of the enzyme molecule, with different states having different kinetic properties. However, it is difficult to understand the underlying mechanism of hysteresis by observing bulk reactions because the different enzyme molecules in the population behave stochastically. In this work, we studied the hysteretic behavior of mutant β-glucuronidase (GUS) using a high-throughput single-molecule array platform and investigated the effect of thermal treatment on the hysteresis.

AB - Hysteresis is an important feature of enzyme-catalyzed reactions, as it reflects the influence of enzyme regulation in the presence of ligands such as substrates or allosteric molecules. In typical kinetic studies of enzyme activity, hysteretic behavior is observed as a "lag" or "burst" in the time course of the catalyzed reaction. These lags and bursts are due to the relatively slow transition from one state to another state of the enzyme molecule, with different states having different kinetic properties. However, it is difficult to understand the underlying mechanism of hysteresis by observing bulk reactions because the different enzyme molecules in the population behave stochastically. In this work, we studied the hysteretic behavior of mutant β-glucuronidase (GUS) using a high-throughput single-molecule array platform and investigated the effect of thermal treatment on the hysteresis.

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Single-Molecule Mechanistic Study of Enzyme Hysteresis

Abstract

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