A Straightforward and Robust Method for Introducing Human Hair as a Nucleant into High Throughput Crystallization Trials

Nederlof, Igor; Hosseini, Rohola; Georgieva, Dilyana; Luo, Jinghui; Li, Dianfan; Abrahams, Jan Pieter

doi:10.1021/cg101374r

Cited by 14 publications

(19 citation statements)

References 31 publications

(47 reference statements)

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“…This have suggested the use of heterogeneous seeding also in initial screening experiments. In fact, recently heterogeneous nucleating agents have been added to a sparse matrix crystallization screen [148] and crystallization plates that are locally coated with fragments of human hair have been prepared [149]. …”

Section: Approaches To Induce Nucleationmentioning

confidence: 99%

An Overview of Biological Macromolecule Crystallization

Krauss

Merlino

Vergara

et al. 2013

IJMS

123

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The elucidation of the three dimensional structure of biological macromolecules has provided an important contribution to our current understanding of many basic mechanisms involved in life processes. This enormous impact largely results from the ability of X-ray crystallography to provide accurate structural details at atomic resolution that are a prerequisite for a deeper insight on the way in which bio-macromolecules interact with each other to build up supramolecular nano-machines capable of performing specialized biological functions. With the advent of high-energy synchrotron sources and the development of sophisticated software to solve X-ray and neutron crystal structures of large molecules, the crystallization step has become even more the bottleneck of a successful structure determination. This review introduces the general aspects of protein crystallization, summarizes conventional and innovative crystallization methods and focuses on the new strategies utilized to improve the success rate of experiments and increase crystal diffraction quality.

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Section: Approaches To Induce Nucleationmentioning

confidence: 99%

An Overview of Biological Macromolecule Crystallization

Krauss

Merlino

Vergara

et al. 2013

IJMS

123

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“…Glucose isomerase (molecular weight = 173 kDa) was crystallized from a solution containing Figure 4(df), significantly larger than that achieved on the substrates S and on several other heterogeneous surfaces 25 . A typical x-ray diffraction pattern for a single glucose isomerase crystal grown on these nano-wrinkled substrates has been presented in Figure 4g for which maximum resolution of 2.5Å could be achieved.…”

Section: Crystallization Of Glucose Isomerase and Xylanase IImentioning

confidence: 88%

Disordered Nanowrinkle Substrates for Inducing Crystallization over a Wide Range of Concentration of Protein and Precipitant

Ghatak

2013

Langmuir

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There are large number of proteins, the existence of which are known but not their crystal structure, because of difficulty in finding the exact condition for their crystallization. Heterogeneous nucleation on disordered porous substrates with small yet large distribution of pores is considered a panacea for this problem, but a universal nucleant suitable for crystallizing large variety of proteins does not really exist. To this end, we report here a nanowrinkled substrate which displays remarkable ability and control over protein crystallization. Experiments with different proteins show that on these substrates crystals nucleate even at very low protein concentration in buffer. A small number of very large crystals appear for precipitant concentrations varied over orders of magnitude, ~0.003-0.3 M; for some proteins, crystals appear even without addition of any precipitant, not seen with any other heterogeneous substrates. In essence, these substrates significantly diminish the influence of the above two parameters, thought to be key factors for crystallization, signifying that this advantage can be exploited for finding out crystallization condition for other yet-to-be-crystallized proteins.

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“…Growing small microcrystals for MicroED is feasible using standard routines, optimizing the crystallization conditions using sitting-drop or hanging-drop vapour diffusion (Georgieva et al, 2007;Nederlof et al, 2011;Calero et al, 2014), and can also be optimized and scaled via batch crystallization (Beale et al, 2019;Wolff et al, 2020). The identification of microcrystals from many different conditions in crystallization plates is not yet straightforward, as small microcrystals fall beyond what can be resolved by optical microscopes, and sometimes it is difficult to distinguish protein crystals from precipitate.…”

Section: Crystallizationmentioning

confidence: 99%

Macromolecular crystallography using microcrystal electron diffraction

Clabbers¹,

Xu²

2021

Acta Crystallogr D Struct Biol

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Microcrystal electron diffraction (MicroED) has recently emerged as a promising method for macromolecular structure determination in structural biology. Since the first protein structure was determined in 2013, the method has been evolving rapidly. Several protein structures have been determined and various studies indicate that MicroED is capable of (i) revealing atomic structures with charges, (ii) solving new protein structures by molecular replacement, (iii) visualizing ligand-binding interactions and (iv) determining membrane-protein structures from microcrystals embedded in lipidic mesophases. However, further development and optimization is required to make MicroED experiments more accurate and more accessible to the structural biology community. Here, we provide an overview of the current status of the field, and highlight the ongoing development, to provide an indication of where the field may be going in the coming years. We anticipate that MicroED will become a robust method for macromolecular structure determination, complementing existing methods in structural biology.

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A Straightforward and Robust Method for Introducing Human Hair as a Nucleant into High Throughput Crystallization Trials

Cited by 14 publications

References 31 publications

An Overview of Biological Macromolecule Crystallization

An Overview of Biological Macromolecule Crystallization

Disordered Nanowrinkle Substrates for Inducing Crystallization over a Wide Range of Concentration of Protein and Precipitant

Macromolecular crystallography using microcrystal electron diffraction

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