Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography Article Swipe
P. Mehrabi
,
Sihyun Sung
,
David von Stetten
,
Andreas Prester
,
Caitlin E. Hatton
,
Stephan Kleine-Döpke
,
Alexander Berkes
,
Gargi Gore
,
Jan-Philipp Leimkohl
,
Hendrik Schikora
,
Martin Kollewe
,
Holger Rohde
,
Matthias Wilmanns
,
Friedjof Tellkamp
,
Eike C. Schulz
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1038/s41467-023-37834-w
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1038/s41467-023-37834-w
We introduce the spitrobot , a protein crystal plunger, enabling reaction quenching via cryo-trapping with a time-resolution in the millisecond range. Protein crystals are mounted on canonical micromeshes on an electropneumatic piston, where the crystals are kept in a humidity and temperature-controlled environment, then reactions are initiated via the liquid application method (LAMA) and plunging into liquid nitrogen is initiated after an electronically set delay time to cryo-trap intermediate states. High-magnification images are automatically recorded before and after droplet deposition, prior to plunging. The SPINE-standard sample holder is directly plunged into a storage puck, enabling compatibility with high-throughput infrastructure. Here we demonstrate binding of glucose and 2,3-butanediol in microcrystals of xylose isomerase, and of avibactam and ampicillin in microcrystals of the extended spectrum beta-lactamase CTX-M-14. We also trap reaction intermediates and conformational changes in macroscopic crystals of tryptophan synthase to demonstrate that the spitrobot enables insight into catalytic events.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1038/s41467-023-37834-w
- https://www.nature.com/articles/s41467-023-37834-w.pdf
- OA Status
- gold
- Cited By
- 20
- References
- 43
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4366988001
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- OpenAlex ID
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https://openalex.org/W4366988001Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1038/s41467-023-37834-wDigital Object Identifier
- Title
-
Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallographyWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2023Year of publication
- Publication date
-
2023-04-25Full publication date if available
- Authors
-
P. Mehrabi, Sihyun Sung, David von Stetten, Andreas Prester, Caitlin E. Hatton, Stephan Kleine-Döpke, Alexander Berkes, Gargi Gore, Jan-Philipp Leimkohl, Hendrik Schikora, Martin Kollewe, Holger Rohde, Matthias Wilmanns, Friedjof Tellkamp, Eike C. SchulzList of authors in order
- Landing page
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https://doi.org/10.1038/s41467-023-37834-wPublisher landing page
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https://www.nature.com/articles/s41467-023-37834-w.pdfDirect link to full text PDF
- Open access
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YesWhether a free full text is available
- OA status
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goldOpen access status per OpenAlex
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https://www.nature.com/articles/s41467-023-37834-w.pdfDirect OA link when available
- Concepts
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Millisecond, Materials science, Trapping, Liquid nitrogen, Microfluidics, Chemistry, Crystallography, Chemical physics, Nanotechnology, Physics, Ecology, Astronomy, Biology, Organic chemistryTop concepts (fields/topics) attached by OpenAlex
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20Total citation count in OpenAlex
- Citations by year (recent)
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2025: 8, 2024: 9, 2023: 3Per-year citation counts (last 5 years)
- References (count)
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43Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.where | 33 |
| abstract_inverted_index.(LAMA) | 53 |
| abstract_inverted_index.before | 76 |
| abstract_inverted_index.holder | 87 |
| abstract_inverted_index.images | 72 |
| abstract_inverted_index.liquid | 50, 57 |
| abstract_inverted_index.method | 52 |
| abstract_inverted_index.range. | 21 |
| abstract_inverted_index.sample | 86 |
| abstract_inverted_index.xylose | 111 |
| abstract_inverted_index.Protein | 22 |
| abstract_inverted_index.binding | 103 |
| abstract_inverted_index.changes | 133 |
| abstract_inverted_index.crystal | 8 |
| abstract_inverted_index.droplet | 79 |
| abstract_inverted_index.enables | 145 |
| abstract_inverted_index.events. | 149 |
| abstract_inverted_index.glucose | 105 |
| abstract_inverted_index.insight | 146 |
| abstract_inverted_index.mounted | 25 |
| abstract_inverted_index.piston, | 32 |
| abstract_inverted_index.plunged | 90 |
| abstract_inverted_index.protein | 7 |
| abstract_inverted_index.states. | 70 |
| abstract_inverted_index.storage | 93 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.crystals | 23, 35, 136 |
| abstract_inverted_index.directly | 89 |
| abstract_inverted_index.enabling | 10, 95 |
| abstract_inverted_index.extended | 122 |
| abstract_inverted_index.humidity | 40 |
| abstract_inverted_index.nitrogen | 58 |
| abstract_inverted_index.plunger, | 9 |
| abstract_inverted_index.plunging | 55 |
| abstract_inverted_index.reaction | 11, 129 |
| abstract_inverted_index.recorded | 75 |
| abstract_inverted_index.spectrum | 123 |
| abstract_inverted_index.synthase | 139 |
| abstract_inverted_index.CTX-M-14. | 125 |
| abstract_inverted_index.avibactam | 115 |
| abstract_inverted_index.canonical | 27 |
| abstract_inverted_index.catalytic | 148 |
| abstract_inverted_index.cryo-trap | 68 |
| abstract_inverted_index.initiated | 47, 60 |
| abstract_inverted_index.introduce | 2 |
| abstract_inverted_index.plunging. | 83 |
| abstract_inverted_index.quenching | 12 |
| abstract_inverted_index.reactions | 45 |
| abstract_inverted_index.spitrobot | 4, 144 |
| abstract_inverted_index.ampicillin | 117 |
| abstract_inverted_index.isomerase, | 112 |
| abstract_inverted_index.tryptophan | 138 |
| abstract_inverted_index.application | 51 |
| abstract_inverted_index.demonstrate | 102, 141 |
| abstract_inverted_index.deposition, | 80 |
| abstract_inverted_index.macroscopic | 135 |
| abstract_inverted_index.micromeshes | 28 |
| abstract_inverted_index.millisecond | 20 |
| abstract_inverted_index.environment, | 43 |
| abstract_inverted_index.intermediate | 69 |
| abstract_inverted_index.automatically | 74 |
| abstract_inverted_index.compatibility | 96 |
| abstract_inverted_index.cryo-trapping | 14 |
| abstract_inverted_index.intermediates | 130 |
| abstract_inverted_index.microcrystals | 109, 119 |
| abstract_inverted_index.2,3-butanediol | 107 |
| abstract_inverted_index.SPINE-standard | 85 |
| abstract_inverted_index.beta-lactamase | 124 |
| abstract_inverted_index.conformational | 132 |
| abstract_inverted_index.electronically | 63 |
| abstract_inverted_index.high-throughput | 98 |
| abstract_inverted_index.infrastructure. | 99 |
| abstract_inverted_index.time-resolution | 17 |
| abstract_inverted_index.electropneumatic | 31 |
| abstract_inverted_index.High-magnification | 71 |
| abstract_inverted_index.temperature-controlled | 42 |
| cited_by_percentile_year.max | 99 |
| cited_by_percentile_year.min | 96 |
| corresponding_author_ids | https://openalex.org/A5032975288 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 15 |
| corresponding_institution_ids | https://openalex.org/I159176309, https://openalex.org/I4210149784 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/9 |
| sustainable_development_goals[0].score | 0.6200000047683716 |
| sustainable_development_goals[0].display_name | Industry, innovation and infrastructure |
| citation_normalized_percentile.value | 0.88389855 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | True |