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Structure of the Lysinibacillus sphaericus Tpp49Aa1 pesticidal protein elucidated from natural crystals using MHz-SFX.

Williamson, Lainey J ; Galchenkova, Marina ; Best, Hannah L ; Bean, Richard J ; Munke, Anna LU orcid ; Awel, Salah ; Pena, Gisel ; Knoska, Juraj ; Schubert, Robin and Dörner, Katerina , et al. (2023) In Proceedings of the National Academy of Sciences of the United States of America 120(49).
Abstract

The
Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito
Culex quinquefasciatus and have potential use in biocontrol. Given that proteins with sequence homology to the individual proteins can have activity alone against other insect species, the structure of Tpp49Aa1 was solved in order to understand this protein more fully and inform the design of improved biopesticides. Tpp49Aa1 is naturally expressed as a crystalline inclusion within the host bacterium, and MHz serial femtosecond crystallography using the novel nanofocus option at an X-ray free electron laser allowed rapid and high-quality data collection to determine the structure of Tpp49Aa1 at 1.62 Å resolution. This... (More)

The
Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito
Culex quinquefasciatus and have potential use in biocontrol. Given that proteins with sequence homology to the individual proteins can have activity alone against other insect species, the structure of Tpp49Aa1 was solved in order to understand this protein more fully and inform the design of improved biopesticides. Tpp49Aa1 is naturally expressed as a crystalline inclusion within the host bacterium, and MHz serial femtosecond crystallography using the novel nanofocus option at an X-ray free electron laser allowed rapid and high-quality data collection to determine the structure of Tpp49Aa1 at 1.62 Å resolution. This revealed the packing of Tpp49Aa1 within these natural nanocrystals as a homodimer with a large intermolecular interface. Complementary experiments conducted at varied pH also enabled investigation of the early structural events leading up to the dissolution of natural Tpp49Aa1 crystals-a crucial step in its mechanism of action. To better understand the cooperation between the two proteins, assays were performed on a range of different mosquito cell lines using both individual proteins and mixtures of the two. Finally, bioassays demonstrated Tpp49Aa1/Cry48Aa1 susceptibility of
Anopheles stephensi,
Aedes albopictus, and
Culex tarsalis larvae-substantially increasing the potential use of this binary toxin in mosquito control.

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Pesticides, Bacillus, Bacillaceae/chemistry, Mosquito Control, Culex, Larva/metabolism
in
Proceedings of the National Academy of Sciences of the United States of America
volume
120
issue
49
article number
e2203241120
publisher
National Academy of Sciences
external identifiers
  • scopus:85178496585
  • pmid:38015839
ISSN
1091-6490
DOI
10.1073/pnas.2203241120
language
English
LU publication?
no
id
25090e81-28cf-470f-acb8-902e9c96684c
date added to LUP
2025-04-21 20:40:12
date last changed
2025-07-01 10:29:43
@article{25090e81-28cf-470f-acb8-902e9c96684c,
  abstract     = {{<p>The <br>
 Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito <br>
 Culex quinquefasciatus and have potential use in biocontrol. Given that proteins with sequence homology to the individual proteins can have activity alone against other insect species, the structure of Tpp49Aa1 was solved in order to understand this protein more fully and inform the design of improved biopesticides. Tpp49Aa1 is naturally expressed as a crystalline inclusion within the host bacterium, and MHz serial femtosecond crystallography using the novel nanofocus option at an X-ray free electron laser allowed rapid and high-quality data collection to determine the structure of Tpp49Aa1 at 1.62 Å resolution. This revealed the packing of Tpp49Aa1 within these natural nanocrystals as a homodimer with a large intermolecular interface. Complementary experiments conducted at varied pH also enabled investigation of the early structural events leading up to the dissolution of natural Tpp49Aa1 crystals-a crucial step in its mechanism of action. To better understand the cooperation between the two proteins, assays were performed on a range of different mosquito cell lines using both individual proteins and mixtures of the two. Finally, bioassays demonstrated Tpp49Aa1/Cry48Aa1 susceptibility of <br>
 Anopheles stephensi, <br>
 Aedes albopictus, and <br>
 Culex tarsalis larvae-substantially increasing the potential use of this binary toxin in mosquito control.<br>
 </p>}},
  author       = {{Williamson, Lainey J and Galchenkova, Marina and Best, Hannah L and Bean, Richard J and Munke, Anna and Awel, Salah and Pena, Gisel and Knoska, Juraj and Schubert, Robin and Dörner, Katerina and Park, Hyun-Woo and Bideshi, Dennis K and Henkel, Alessandra and Kremling, Viviane and Klopprogge, Bjarne and Lloyd-Evans, Emyr and Young, Mark T and Valerio, Joana and Kloos, Marco and Sikorski, Marcin and Mills, Grant and Bielecki, Johan and Kirkwood, Henry and Kim, Chan and de Wijn, Raphael and Lorenzen, Kristina and Xavier, Paul Lourdu and Rahmani Mashhour, Aida and Gelisio, Luca and Yefanov, Oleksandr and Mancuso, Adrian P and Federici, Brian A and Chapman, Henry N and Crickmore, Neil and Rizkallah, Pierre J and Berry, Colin and Oberthür, Dominik}},
  issn         = {{1091-6490}},
  keywords     = {{Animals; Pesticides; Bacillus; Bacillaceae/chemistry; Mosquito Control; Culex; Larva/metabolism}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{49}},
  publisher    = {{National Academy of Sciences}},
  series       = {{Proceedings of the National Academy of Sciences of the United States of America}},
  title        = {{Structure of the <i>Lysinibacillus sphaericus </i>Tpp49Aa1 pesticidal protein elucidated from natural crystals using MHz-SFX.}},
  url          = {{http://dx.doi.org/10.1073/pnas.2203241120}},
  doi          = {{10.1073/pnas.2203241120}},
  volume       = {{120}},
  year         = {{2023}},
}