---
_id: '9205'
abstract:
- lang: eng
  text: Cryo-EM grid preparation is an important bottleneck in protein structure determination,
    especially for membrane proteins, typically requiring screening of a large number
    of conditions. We systematically investigated the effects of buffer components,
    blotting conditions and grid types on the outcome of grid preparation of five
    different membrane protein samples. Aggregation was the most common type of problem
    which was addressed by changing detergents, salt concentration or reconstitution
    of proteins into nanodiscs or amphipols. We show that the optimal concentration
    of detergent is between 0.05 and 0.4% and that the presence of a low concentration
    of detergent with a high critical micellar concentration protects the proteins
    from denaturation at the air-water interface. Furthermore, we discuss the strategies
    for achieving an adequate ice thickness, particle coverage and orientation distribution
    on free ice and on support films. Our findings provide a clear roadmap for comprehensive
    screening of conditions for cryo-EM grid preparation of membrane proteins.
acknowledged_ssus:
- _id: EM-Fac
acknowledgement: We thank the Electron Microscopy Facilities at the Institute of Science
  and Technology Austria and at the Vienna Biocenter for providing access and training
  for the electron microscopes. This project has received funding from the European
  Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie
  Grant Agreement no. 665385 .
article_number: '102139'
article_processing_charge: No
article_type: original
author:
- first_name: Domen
  full_name: Kampjut, Domen
  id: 37233050-F248-11E8-B48F-1D18A9856A87
  last_name: Kampjut
- first_name: Julia
  full_name: Steiner, Julia
  id: 3BB67EB0-F248-11E8-B48F-1D18A9856A87
  last_name: Steiner
- first_name: Leonid A
  full_name: Sazanov, Leonid A
  id: 338D39FE-F248-11E8-B48F-1D18A9856A87
  last_name: Sazanov
  orcid: 0000-0002-0977-7989
citation:
  ama: Kampjut D, Steiner J, Sazanov LA. Cryo-EM grid optimization for membrane proteins.
    <i>iScience</i>. 2021;24(3). doi:<a href="https://doi.org/10.1016/j.isci.2021.102139">10.1016/j.isci.2021.102139</a>
  apa: Kampjut, D., Steiner, J., &#38; Sazanov, L. A. (2021). Cryo-EM grid optimization
    for membrane proteins. <i>IScience</i>. Elsevier. <a href="https://doi.org/10.1016/j.isci.2021.102139">https://doi.org/10.1016/j.isci.2021.102139</a>
  chicago: Kampjut, Domen, Julia Steiner, and Leonid A Sazanov. “Cryo-EM Grid Optimization
    for Membrane Proteins.” <i>IScience</i>. Elsevier, 2021. <a href="https://doi.org/10.1016/j.isci.2021.102139">https://doi.org/10.1016/j.isci.2021.102139</a>.
  ieee: D. Kampjut, J. Steiner, and L. A. Sazanov, “Cryo-EM grid optimization for
    membrane proteins,” <i>iScience</i>, vol. 24, no. 3. Elsevier, 2021.
  ista: Kampjut D, Steiner J, Sazanov LA. 2021. Cryo-EM grid optimization for membrane
    proteins. iScience. 24(3), 102139.
  mla: Kampjut, Domen, et al. “Cryo-EM Grid Optimization for Membrane Proteins.” <i>IScience</i>,
    vol. 24, no. 3, 102139, Elsevier, 2021, doi:<a href="https://doi.org/10.1016/j.isci.2021.102139">10.1016/j.isci.2021.102139</a>.
  short: D. Kampjut, J. Steiner, L.A. Sazanov, IScience 24 (2021).
date_created: 2021-02-28T23:01:24Z
date_published: 2021-03-19T00:00:00Z
date_updated: 2023-08-07T13:54:06Z
day: '19'
ddc:
- '570'
department:
- _id: LeSa
doi: 10.1016/j.isci.2021.102139
ec_funded: 1
external_id:
  isi:
  - '000631646000012'
  pmid:
  - '33665558'
file:
- access_level: open_access
  checksum: 50585447386fe5842f07ab9b3a66e7e9
  content_type: application/pdf
  creator: dernst
  date_created: 2021-03-03T07:38:14Z
  date_updated: 2021-03-03T07:38:14Z
  file_id: '9219'
  file_name: 2021_iScience_Kampjut.pdf
  file_size: 7431411
  relation: main_file
  success: 1
file_date_updated: 2021-03-03T07:38:14Z
has_accepted_license: '1'
intvolume: '        24'
isi: 1
issue: '3'
language:
- iso: eng
month: '03'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 2564DBCA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '665385'
  name: International IST Doctoral Program
publication: iScience
publication_identifier:
  eissn:
  - '25890042'
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Cryo-EM grid optimization for membrane proteins
tmp:
  image: /images/cc_by_nc_nd.png
  legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
  name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
    (CC BY-NC-ND 4.0)
  short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 24
year: '2021'
...