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Cited by 3Protein Structure and DynamicsEvolution and Genetic DynamicsBacterial Genetics and Biotechnology

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Evolutionary Engineering a Larger Porin Using a Loop-to-Hairpin Mechanism

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Dhar, Rik, Bowman, Alexander M., Hatungimana, Brunojoel, SG Slusky, Joanna·2023·Journal of Molecular Biology
DOI 10.1016/j.jmb.2023.168292PMC PMC11215794

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Abstract

In protein evolution, diversification is generally driven by genetic duplication. The hallmarks of this mechanism are visible in the repeating topology of various proteins. In outer membrane β-barrels, duplication is visible with β-hairpins as the repeating unit of the barrel. In contrast to the overall use of duplication in diversification, a computational study hypothesized evolutionary mechanisms other than hairpin duplications leading to increases in the number of strands in outer membrane β-barrels. Specifically, the topology of some 16- and 18-stranded β-barrels appear to have evolved through a loop to β-hairpin transition. Here we test this novel evolutionary mechanism by creating a chimeric protein from an 18-stranded β-barrel and an evolutionarily related 16-stranded β-barrel. The chimeric combination of the two was created by replacing loop L3 of the 16-stranded barrel with the sequentially matched transmembrane β-hairpin region of the 18-stranded barrel. We find the resulting chimeric protein is stable and has characteristics of increased strand number. This study provides the first experimental evidence supporting the evolution through a loop to β-hairpin transition. Highlights: We find evidence supporting a novel diversification mechanism in membrane β-barrelsThe mechanism is the conversion of an extracellular loop to transmembrane β-hairpinA chimeric protein modeling this mechanism folds stably in the membraneThe chimera has more β-structure and a larger pore, consistent with a loop-to-hairpin transition.

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Metadata

Authors
Dhar, R., Bowman, A. M., Hatungimana, B., & SG Slusky, J.
Affiliations
University of Kansas (US)
ORCID
Venue
Journal of Molecular Biology
Publisher
Elsevier BV
Published
2023-11
Volume
435
Issue
22
Pages
168292
ISSN
0022-2836 (ISSN-L)
DOI
10.1016/j.jmb.2023.168292
PMC
PMC11215794
OpenAlex
W4387046787
Cited by
3
References
1
Citations by year
2026: 2 · 2024: 1
FWCI
0.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Molecular Biology (publishedVersion, cc-by-nc)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • PubMed Central (submittedVersion, cc-by-nc-nd)
Funders
  • National Science Foundation (2226804)
  • National Institute of General Medical Sciences (DP2GM128201, P20GM103418, P20GM113117, R01GM148583)
  • National Science Foundation Directorate for Biological Sciences
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Evolution and Genetic Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
porin, protein evolution, fusion protein, beta-hairpin, protein stability, protein topology

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CoChat. (2026). Citation check: Evolutionary Engineering a Larger Porin Using a Loop-to-Hairpin Mechanism. https://app.cochat.ai/papers/10.1016/j.jmb.2023.168292
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Dhar, R., Bowman, A. M., Hatungimana, B., & SG Slusky, J. (2023). Evolutionary Engineering a Larger Porin Using a Loop-to-Hairpin Mechanism. Journal of Molecular Biology. https://doi.org/10.1016/j.jmb.2023.168292