De novo designed proteins from a library of artificial sequences function in Escherichia coli and enable cell growth
A central challenge of synthetic biology is to enable the growth of living systems using parts that are not derived from nature, but designed and synthesized in the laboratory. As an initial step toward achieving this goal, we probed the ability of a collection of >10(6) de novo designed proteins...
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Published in | PloS one Vol. 6; no. 1; p. e15364 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Public Library of Science
04.01.2011
Public Library of Science (PLoS) |
Subjects | |
Online Access | Get full text |
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Summary: | A central challenge of synthetic biology is to enable the growth of living systems using parts that are not derived from nature, but designed and synthesized in the laboratory. As an initial step toward achieving this goal, we probed the ability of a collection of >10(6) de novo designed proteins to provide biological functions necessary to sustain cell growth. Our collection of proteins was drawn from a combinatorial library of 102-residue sequences, designed by binary patterning of polar and nonpolar residues to fold into stable 4-helix bundles. We probed the capacity of proteins from this library to function in vivo by testing their abilities to rescue 27 different knockout strains of Escherichia coli, each deleted for a conditionally essential gene. Four different strains--ΔserB, ΔgltA, ΔilvA, and Δfes--were rescued by specific sequences from our library. Further experiments demonstrated that a strain simultaneously deleted for all four genes was rescued by co-expression of four novel sequences. Thus, cells deleted for ∼0.1% of the E. coli genome (and ∼1% of the genes required for growth under nutrient-poor conditions) can be sustained by sequences designed de novo. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1 Conceived and designed the experiments: MAF KLM LHB SRV MHH. Performed the experiments: MAF KLM LHB SRV. Analyzed the data: MAF KLM LHB SRV MHH. Contributed reagents/materials/analysis tools: MAF KLM LHB SRV MHH. Wrote the paper: MAF KLM LHB SRV MHH. Current address: Columbia University Medical Center, New York, New York, United States of America Current address: Departments of Anatomy and Neurobiology, Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America Current address: Energy Biosciences Institute, University of California, Berkeley, California, United States of America Current address: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America |
ISSN: | 1932-6203 1932-6203 |
DOI: | 10.1371/journal.pone.0015364 |