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Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography

Standard

Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography. / Mehrabi, Pedram; Sung, Sihyun; von Stetten, David; Prester, Andreas; Hatton, Caitlin E; Kleine-Döpke, Stephan; Berkes, Alexander; Gore, Gargi; Leimkohl, Jan-Philipp; Schikora, Hendrik; Kollewe, Martin; Rohde, Holger; Wilmanns, Matthias; Tellkamp, Friedjof; Schulz, Eike C.

In: NAT COMMUN, Vol. 14, No. 1, 25.04.2023, p. 2365.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Mehrabi, P, Sung, S, von Stetten, D, Prester, A, Hatton, CE, Kleine-Döpke, S, Berkes, A, Gore, G, Leimkohl, J-P, Schikora, H, Kollewe, M, Rohde, H, Wilmanns, M, Tellkamp, F & Schulz, EC 2023, 'Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography', NAT COMMUN, vol. 14, no. 1, pp. 2365. https://doi.org/10.1038/s41467-023-37834-w

APA

Mehrabi, P., Sung, S., von Stetten, D., Prester, A., Hatton, C. E., Kleine-Döpke, S., Berkes, A., Gore, G., Leimkohl, J-P., Schikora, H., Kollewe, M., Rohde, H., Wilmanns, M., Tellkamp, F., & Schulz, E. C. (2023). Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography. NAT COMMUN, 14(1), 2365. https://doi.org/10.1038/s41467-023-37834-w

Vancouver

Mehrabi P, Sung S, von Stetten D, Prester A, Hatton CE, Kleine-Döpke S et al. Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography. NAT COMMUN. 2023 Apr 25;14(1):2365. https://doi.org/10.1038/s41467-023-37834-w

Bibtex

@article{d8f9a608efa648b2b0d1d0fead31d4be,
title = "Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography",
abstract = "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.",
keywords = "Crystallography/methods, Proteins/chemistry, Temperature, Humidity, Crystallography, X-Ray",
author = "Pedram Mehrabi and Sihyun Sung and {von Stetten}, David and Andreas Prester and Hatton, {Caitlin E} and Stephan Kleine-D{\"o}pke and Alexander Berkes and Gargi Gore and Jan-Philipp Leimkohl and Hendrik Schikora and Martin Kollewe and Holger Rohde and Matthias Wilmanns and Friedjof Tellkamp and Schulz, {Eike C}",
note = "{\textcopyright} 2023. The Author(s).",
year = "2023",
month = apr,
day = "25",
doi = "10.1038/s41467-023-37834-w",
language = "English",
volume = "14",
pages = "2365",
journal = "NAT COMMUN",
issn = "2041-1723",
publisher = "NATURE PUBLISHING GROUP",
number = "1",

}

RIS

TY - JOUR

T1 - Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography

AU - Mehrabi, Pedram

AU - Sung, Sihyun

AU - von Stetten, David

AU - Prester, Andreas

AU - Hatton, Caitlin E

AU - Kleine-Döpke, Stephan

AU - Berkes, Alexander

AU - Gore, Gargi

AU - Leimkohl, Jan-Philipp

AU - Schikora, Hendrik

AU - Kollewe, Martin

AU - Rohde, Holger

AU - Wilmanns, Matthias

AU - Tellkamp, Friedjof

AU - Schulz, Eike C

N1 - © 2023. The Author(s).

PY - 2023/4/25

Y1 - 2023/4/25

N2 - 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.

AB - 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.

KW - Crystallography/methods

KW - Proteins/chemistry

KW - Temperature

KW - Humidity

KW - Crystallography, X-Ray

U2 - 10.1038/s41467-023-37834-w

DO - 10.1038/s41467-023-37834-w

M3 - SCORING: Journal article

C2 - 37185266

VL - 14

SP - 2365

JO - NAT COMMUN

JF - NAT COMMUN

SN - 2041-1723

IS - 1

ER -