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Measuring gas vesicle dimensions by electron microscopy

Dutka, Przemysław and Malounda, Dina and Metskas, Lauren Ann and Chen, Songye and Hurt, Robert C. and Lu, George J. and Jensen, Grant J. and Shapiro, Mikhail G. (2021) Measuring gas vesicle dimensions by electron microscopy. Protein Science, 30 (5). pp. 1081-1086. ISSN 0961-8368. PMCID PMC8040859. doi:10.1002/pro.4056. https://resolver.caltech.edu/CaltechAUTHORS:20210125-082358436

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Abstract

Gas vesicles (GVs) are cylindrical or spindle‐shaped protein nanostructures filled with air and used for flotation by various cyanobacteria, heterotrophic bacteria, and Archaea. Recently, GVs have gained interest in biotechnology applications due to their ability to serve as imaging agents and actuators for ultrasound, magnetic resonance and several optical techniques. The diameter of GVs is a crucial parameter contributing to their mechanical stability, buoyancy function and evolution in host cells, as well as their properties in imaging applications. Despite its importance, reported diameters for the same types of GV differ depending on the method used for its assessment. Here, we provide an explanation for these discrepancies and utilize electron microscopy (EM) techniques to accurately estimate the diameter of the most commonly studied types of GVs. We show that during air drying on the EM grid, GVs flatten, leading to a ~1.5‐fold increase in their apparent diameter. We demonstrate that GVs' diameter can be accurately determined by direct measurements from cryo‐EM samples or alternatively indirectly derived from widths of flat collapsed and negatively stained GVs. Our findings help explain the inconsistency in previously reported data and provide accurate methods to measure GVs dimensions.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/pro.4056DOIArticle
https://doi.org/10.1101/2021.01.22.427725DOIDiscussion Paper
ORCID:
AuthorORCID
Dutka, Przemysław0000-0003-3819-1618
Malounda, Dina0000-0001-7086-9877
Metskas, Lauren Ann0000-0002-8073-6960
Chen, Songye0000-0001-5407-5049
Hurt, Robert C.0000-0002-4347-6901
Lu, George J.0000-0002-4689-9686
Jensen, Grant J.0000-0003-1556-4864
Shapiro, Mikhail G.0000-0002-0291-4215
Additional Information:© 2021 The Protein Society. Issue Online: 12 April 2021; Version of Record online: 11 March 2021; Accepted manuscript online: 27 February 2021; Manuscript accepted: 22 February 2021; Manuscript received: 22 January 2021. This work was supported by the National Institutes of Health (grant R35-GM122588 to G.J.J. and R01-EB018975 to M.G.S.) and the Caltech Center for Environmental Microbial Interactions (CEMI). Electron microscopy was performed in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. Related research in the Shapiro Laboratory is also supported by the Heritage Medical Research Institute, the Pew Scholarship in the Biomedical Sciences, and the Packard Fellowship for Science and Engineering. Author Contributions: Przemysław Dutka: Conceptualization; methodology; investigation; formal analysis; visualization; writing – original draft preparation; writing – review editing. Dina Malounda: Investigation. Lauren Ann Metskas: Investigation. Songye Chen: Investigation. Robert C. Hurt: Investigation. George J. Lu: Investigation. Grant J. Jensen: Conceptualization; writing – review editing; supervision; funding acquisition. Mikhail G. Shapiro: Conceptualization; writing – review editing; supervision; funding acquisition. The authors declare no competing interests.
Group:Caltech Center for Environmental Microbial Interactions (CEMI), Heritage Medical Research Institute
Funders:
Funding AgencyGrant Number
NIHR35-GM122588
NIHR01-EB018975
Caltech Center for Environmental Microbial Interactions (CEMI)UNSPECIFIED
Heritage Medical Research InstituteUNSPECIFIED
Pew Charitable TrustUNSPECIFIED
David and Lucile Packard FoundationUNSPECIFIED
Subject Keywords:critical collapse pressure; cryo‐electron microscopy; diameter; electron microscopy; gas vesicles; negative staining
Issue or Number:5
PubMed Central ID:PMC8040859
DOI:10.1002/pro.4056
Record Number:CaltechAUTHORS:20210125-082358436
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210125-082358436
Official Citation:Dutka, P, Malounda, D, Metskas, LA, et al. Measuring gas vesicle dimensions by electron microscopy. Protein Science. 2021; 30: 1081–1086. https://doi.org/10.1002/pro.4056
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107703
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:26 Jan 2021 20:18
Last Modified:04 Oct 2021 22:19

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