Large-scale analysis of macromolecular crowding effects on protein aggregation using a reconstituted cell-free translation system.
Proteins must fold into their native structures in the crowded cellular environment, to perform their functions. Although such macromolecular crowding has been considered to affect the folding properties of proteins, large-scale experimental data have so far been lacking. Here, we individually translated 142 Escherichia coli cytoplasmic proteins using a reconstituted cell-free translation system in the presence of macromolecular crowding reagents (MCRs), Ficoll 70 or dextran 70, and evaluated the aggregation propensities of 142 proteins. The results showed that the MCR effects varied depending on the proteins, although the degree of these effects was modest. Statistical analyses suggested that structural parameters were involved in the effects of the MCRs. Our dataset provides a valuable resource to understand protein folding and aggregation inside cells.
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology Yokohama, Japan.
Tatsuya
T
Niwa
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology Yokohama, Japan.
Ryota
R
Sugimoto
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology Yokohama, Japan.
Lisa
L
Watanabe
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo Tokyo, Japan.
Shugo
S
Nakamura
Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo Chiba, Japan.
Takuya
T
Ueda
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology Yokohama, Japan.
Hideki
H
Taguchi
cell-free translation system; large-scale analysis; macromolecular crowding; protein aggregation; protein folding