Gentle, fast and effective crystal soaking by acoustic dispensing

Collins, P.; Ng, Jia Tsing; Talon, R.; Nekrosiute, Karolina; Krojer, T.; Douangamath, A.; Brandão-Neto, J.; Wright, Nathan; Pearce, Nicholas M.; Delft, Frank von

doi:10.1107/s205979831700331x

Cited by 81 publications

(76 citation statements)

References 35 publications

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“…Soaking crystals when many compounds are to be evaluated might be a tedious process, as the addition of the concentrated ligand solution on the crystal or the transfer of each crystal in a concentrated droplet of ligand is manually performed. However, automation of the soaking procedure has recently been developed using for instance acoustic dispensing of nanodrops of concentrated ligands into crystal containing droplets, where several hundreds of soaking experiments can be performed within minutes [34]. Table 1.…”

Section: Soakingmentioning

confidence: 99%

Protein X-ray Crystallography and Drug Discovery

2020

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With the advent of structural biology in the drug discovery process, medicinal chemists gained the opportunity to use detailed structural information in order to progress screening hits into leads or drug candidates. X-ray crystallography has proven to be an invaluable tool in this respect, as it is able to provide exquisitely comprehensive structural information about the interaction of a ligand with a pharmacological target. As fragment-based drug discovery emerged in the recent years, X-ray crystallography has also become a powerful screening technology, able to provide structural information on complexes involving low-molecular weight compounds, despite weak binding affinities. Given the low numbers of compounds needed in a fragment library, compared to the hundreds of thousand usually present in drug-like compound libraries, it now becomes feasible to screen a whole fragment library using X-ray crystallography, providing a wealth of structural details that will fuel the fragment to drug process. Here, we review theoretical and practical aspects as well as the pros and cons of using X-ray crystallography in the drug discovery process.

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Section: Soakingmentioning

confidence: 99%

Protein X-ray Crystallography and Drug Discovery

2020

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“…In ADE high frequency sound waves are produced by a transducer, which propagates through the liquid medium and ejects a train of droplets at the surface with great accuracy in volume (nanoliter to picoliter), trajectory and frequency [149]. Being non-contact ADE has distinct advantages in liquid handling and has found great application throughout the crystallography pipeline starting from crystallization [150], microseeding [151], and ligand soaking [152][153][154] to crystal mounting [155,156]. Since droplets are ejected at a frequency compatible with the repetition rate of first generation XFEL facilities, ADE has also been utilized as a sample delivery method at both XFEL and synchrotron [157][158][159][160].…”

Section: Drop-on-demand-potential To Maximize Sample Efficiencymentioning

confidence: 99%

Towards an Optimal Sample Delivery Method for Serial Crystallography at XFEL

Cheng¹

2020

Crystals

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The advent of the X-ray free electron laser (XFEL) in the last decade created the discipline of serial crystallography but also the challenge of how crystal samples are delivered to X-ray. Early sample delivery methods demonstrated the proof-of-concept for serial crystallography and XFEL but were beset with challenges of high sample consumption, jet clogging and low data collection efficiency. The potential of XFEL and serial crystallography as the next frontier of structural solution by X-ray for small and weakly diffracting crystals and provision of ultra-fast time-resolved structural data spawned a huge amount of scientific interest and innovation. To utilize the full potential of XFEL and broaden its applicability to a larger variety of biological samples, researchers are challenged to develop better sample delivery methods. Thus, sample delivery is one of the key areas of research and development in the serial crystallography scientific community. Sample delivery currently falls into three main systems: jet-based methods, fixed-target chips, and drop-on-demand. Huge strides have since been made in reducing sample consumption and improving data collection efficiency, thus enabling the use of XFEL for many biological systems to provide high-resolution, radiation damage-free structural data as well as time-resolved dynamics studies. This review summarizes the current main strategies in sample delivery and their respective pros and cons, as well as some future direction.

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“…Te nf urthers caffolds were prepared by using Ugi reactions (Scheme 1). [13] The required cyclisation precursors (19,22,25 and 28)w ere generally prepared by ring opening of ac yclic sulfamidate 6 with ap ropargylic sulfonamide 5.T he outcome of the reaction of these cyclisation precursors with 5mol % Au(PPh 3 )Cl and 5mol %A gSbF 6 in dioxane at 100 8Cd epended on substitution. [13a] Thus, with the terminala lkynes, 19, 22 and 25,t he Boc-protected amine cyclisedo nto promixal alkyne carbon to yield the corresponding tetrahydropyrazines 20, 23 and 26.I nc ontrast, the 1,2-disubstituted alkyne 28 underwent hydration at the remote alkyne carbon to yield the open-chain b-keto amine derivative 29.T he related b-keto amine derivative 31 was also prepared by conjugate addition of the amine 8 to the enone 7.…”

Section: Acquisition Of Ashape-diverse Fragments Etmentioning

confidence: 99%

Construction of a Shape‐Diverse Fragment Set: Design, Synthesis and Screen against Aurora‐A Kinase

Zhang

McIntyre

Collins

et al. 2019

Chemistry A European J

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Historically,c hemists have explored chemical space in ah ighly unevena nd unsystematic manner.A sa n example, the shape diversity of existing fragments ets does not generally reflect that of all theoretically possible fragments. To assess experimentallyt he added value of increased three dimensionality,ashape-diverse fragment set was designed and collated. The set was assembled by both using commerciallya vailable fragments and harnessing unified synthetic approaches to sp 3 -rich molecular scaffolds. The resulting set of 80 fragments was highly three-dimen-sional, and its shape diversity was significantly enriched by twenty synthesised fragments. The fragment set was screened by high-throughput protein crystallography against Aurora-A kinase, revealing four hits that targeted the binding site of allosteric regulators.I nt he longer term, it is envisaged that the fragment set could be screened against a range of functionally diverse proteins, allowing the added value of more shape-diverse screening collections to be more fully assessed.[a] Dr.(South Africa)[g] Prof. R. Bayliss SchoolofM olecular and Cellular Biology,U niversity of Leeds Leeds, LS2 9JT (UK)Supporting information and the ORCID identification number(s) for the author(s) of this articlecan be found under: https://doi.

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Gentle, fast and effective crystal soaking by acoustic dispensing

Abstract: A high-throughput method is described for crystal soaking using acoustic droplet ejection, and its effectiveness is demonstrated.

Cited by 81 publications

References 35 publications

Protein X-ray Crystallography and Drug Discovery

Protein X-ray Crystallography and Drug Discovery

Towards an Optimal Sample Delivery Method for Serial Crystallography at XFEL

Construction of a Shape‐Diverse Fragment Set: Design, Synthesis and Screen against Aurora‐A Kinase

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