气候变化领域集成服务门户(CCPortal): Designing heterotropically activated allosteric conformational switches using supercharging

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DOI	10.1073/pnas.1916046117
	Designing heterotropically activated allosteric conformational switches using supercharging
	Schnatz P.J.; Brisendine J.M.; Laing C.C.; Everson B.H.; French C.A.; Molinaro P.M.; Koder R.L.
发表日期	2020
ISSN	0027-8424
起始页码	5291
结束页码	5297
卷号	117 期号:10
英文摘要	Heterotropic allosteric activation of protein function, in which binding of one ligand thermodynamically activates the binding of another, different ligand or substrate, is a fundamental control mechanism in metabolism and as such has been a long-aspired capability in protein design. Here we show that greatly increasing the magnitude of a protein’s net charge using surface supercharging transforms that protein into an allosteric ligand- and counterion-gated conformational molecular switch. To demonstrate this we first modified the designed helical bundle hemoprotein H4, creating a highly charged protein which both unfolds reversibly at low ionic strength and undergoes the ligand-induced folding transition commonly observed in signal transduction by intrinsically disordered proteins in biology. As a result of the high surface-charge density, ligand binding to this protein is allosterically activated up to 1,300-fold by low concentrations of divalent cations and the polyamine spermine. To extend this process further using a natural protein, we similarly modified Escherichia coli cytochrome b562 and the resulting protein behaves in a like manner. These simple model systems not only establish a set of general engineering principles which can be used to convert natural and designed soluble proteins into allosteric molecular switches useful in biodesign, sensing, and synthetic biology, the behavior we have demonstrated––functional activation of supercharged intrinsically disordered proteins by low concentrations of multivalent ions––may be a control mechanism utilized by Nature which has yet to be appreciated. © 2020 National Academy of Sciences. All rights reserved.
英文关键词	Allostery; Intrinsically disordered proteins; Ligand-induced folding; Protein design; Supercharging
语种	英语
scopus关键词	cytochrome b; divalent cation; hemoprotein; hemoprotein H4; intrinsically disordered protein; sodium chloride; spermine; unclassified drug; calcium; cytochrome b562, E coli; Escherichia coli protein; ligand; magnesium; allosteric conformational switch; allosterism; Article; controlled study; Escherichia coli; ionic strength; ligand binding; nonhuman; priority journal; protein conformation; protein engineering; protein folding; protein structure; protein unfolding; signal transduction; surface charge; chemistry; procedures; thermodynamics; Allosteric Regulation; Calcium; Cations, Divalent; Cytochrome b Group; Escherichia coli Proteins; Hemeproteins; Intrinsically Disordered Proteins; Ligands; Magnesium; Protein Conformation; Protein Engineering; Protein Folding; Spermine; Thermodynamics
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160309
作者单位	Schnatz, P.J., Department of Physics, City College of New York, New York, NY 10031, United States; Brisendine, J.M., Department of Biochemistry, City College of New York, New York, NY 10031, United States; Laing, C.C., Department of Physics, City College of New York, New York, NY 10031, United States; Everson, B.H., Department of Physics, City College of New York, New York, NY 10031, United States; French, C.A., Department of Biochemistry, City College of New York, New York, NY 10031, United States; Molinaro, P.M., Department of Physics, City College of New York, New York, NY 10031, United States; Koder, R.L., Department of Physics, City College of New York, New York, NY 10031, United States, Graduate Program of Physics, Graduate Center, City University of New York, New York, NY 10016, United States, Graduate Program of Biology, Graduate Center, City University of New York, New York, NY 10016, United States, Graduate Program of Biochemistry, Graduate Center, City University of New York, New York,...
推荐引用方式 GB/T 7714	Schnatz P.J.,Brisendine J.M.,Laing C.C.,et al. Designing heterotropically activated allosteric conformational switches using supercharging[J],2020,117(10).
APA	Schnatz P.J..,Brisendine J.M..,Laing C.C..,Everson B.H..,French C.A..,...&Koder R.L..(2020).Designing heterotropically activated allosteric conformational switches using supercharging.Proceedings of the National Academy of Sciences of the United States of America,117(10).
MLA	Schnatz P.J.,et al."Designing heterotropically activated allosteric conformational switches using supercharging".Proceedings of the National Academy of Sciences of the United States of America 117.10(2020).