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Cell‐Like Nanostructured Environments Alter Diffusion and Reaction Kinetics in Cell‐Free Gene Expression
Author(s) -
Hansen Maike M. K.,
Paffenholz Sabine,
Foschepoth David,
Heus Hans A.,
Thiele Julian,
Huck Wilhelm T. S.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500560
Subject(s) - macromolecular crowding , intracellular , cytoplasm , gene expression , macromolecule , kinetics , biophysics , translation (biology) , transcription (linguistics) , microbiology and biotechnology , crowding , cell , chemistry , biology , gene , messenger rna , biochemistry , physics , linguistics , philosophy , quantum mechanics , neuroscience
In highly crowded and viscous intracellular environments, the kinetics of complex enzymatic reactions are determined by both reaction and diffusion rates. However in vitro studies on transcription and translation often fail to take into account the density of the prokaryotic cytoplasm. Here we mimic the cellular environment by using a porous hydrogel matrix, to study the effects of macromolecular crowding on gene expression. We found that within microgels gene expression is localized, transcription is enhanced up to fivefold, and translation is enhanced up to fourfold. Our results highlight the need to consider the role of the physical environment on complex biochemical reactions, in this case macromolecular crowding, nanoscale spatial organization, and confinement.