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  Aerosol-based ligand soaking of reservoir-free protein crystals

Ross, B., Krapp, S., Geiss-Friedlander, R., Littmann, W., Huber, R., & Kiefersauer, R. (2021). Aerosol-based ligand soaking of reservoir-free protein crystals. Journal of Applied Crystallography, 54, 895-902. doi:10.1107/S1600576721003551.

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Genre: Journal Article

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 Creators:
Ross, Breyan1, Author              
Krapp, Stephan2, Author
Geiss-Friedlander, Ruth2, Author
Littmann, Walter2, Author
Huber, Robert1, Author
Kiefersauer, Reiner1, Author              
Affiliations:
1Huber, Robert / Structure Research, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565155              
2external, ou_persistent22              

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Free keywords: DIPEPTIDYL PEPTIDASES 8; SUCCESSFUL GENERATION; SOLUBILITY; POTENT; SYSTEMChemistry; Crystallography; aerosols; soaking; ultrasonic; DPP8;
 Abstract: Soaking of macromolecular crystals allows the formation of complexes via diffusion of molecules into a preformed crystal for structural analysis. Soaking offers various advantages over co-crystallization, e.g. small samples and high-throughput experimentation. However, this method has disadvantages, such as inducing mechanical stress on crystals and reduced success rate caused by low affinity/solubility of the ligand. To bypass these issues, the Picodropper was previously developed in the authors' laboratory. This technique aimed to deliver small volumes of compound solution in response to crystal dehydration supported by the Free Mounting System humidity control or by IR-laser-induced protein crystal transformation. Herein, a new related soaking development, the Aerosol-Generator, is introduced. This device delivers compounds onto the solution-free surface of protein crystals using an ultrasonic technique. The produced aerosol stream enables an easier and more accurate control of solution volumes, reduced crystal handling, and crystal-size-independent soaking. The Aerosol-Generator has been used to produce complexes of DPP8 crystals, where otherwise regular soaking did not achieve complex formation. These results demonstrate the potential of this device in challenging ligand-binding scenarios and contribute to further understanding of DPP8 inhibitor binding.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published in print
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000659339200018
DOI: 10.1107/S1600576721003551
 Degree: -

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Title: Journal of Applied Crystallography
  Abbreviation : J. Appl. Cryst.
Source Genre: Journal
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Publ. Info: Oxford, England : Blackwell Publishing on behalf of the International Union of Crystallography
Pages: - Volume / Issue: 54 Sequence Number: - Start / End Page: 895 - 902 Identifier: ISSN: 0021-8898
CoNE: https://pure.mpg.de/cone/journals/resource/954925410812