Investigation of the Initial Crystallization Stage in Lysozyme Solutions by Small-Angle X-ray Scattering

Kovalchuk, M. V.; Благов, А. Е.; Dyakova, Yulia A.; Gruzinov, Andrey; Марченкова, М. А.; Peters, G. S.; Pisarevsky, Yury V.; Тимофеев, В. И.; Волков, В. В.

doi:10.1021/acs.cgd.5b01662

Cited by 56 publications

(44 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…SAXS profiles provide information on size, shape and oligomerization state but also about interactions between particles in solution. SAXS is extremely sensitive to the formation of crystallites, and this technique has previously been used to analyse protein nucleation (Kovalchuk et al, 2016) and crystallization kinetics (Poplewska et al, 2019). Furthermore, the micro-and nano-GISAX method could even significantly exceed the sensitivity of the SAXS technique for studying protein nucleation (Pechkova & Nicolini, 2017).…”

Section: Introductionmentioning

confidence: 99%

Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

et al. 2020

View full text Add to dashboard Cite

Crystallization of recombinant proteins in living cells is an exciting new approach for structural biology that provides an alternative to the time-consuming optimization of protein purification and extensive crystal screening steps. Exploiting the potential of this approach requires a more detailed understanding of the cellular processes involved and versatile screening strategies for crystals in a cell culture. Particularly if the target protein forms crystalline structures of unknown morphology only in a small fraction of cells, their detection by applying standard visualization techniques can be time consuming and difficult owing to the environmental challenges imposed by the living cells. In this study, a high-brilliance and low-background bioSAXS beamline is employed for rapid and sensitive detection of protein microcrystals grown within insect cells. On the basis of the presence of Bragg peaks in the recorded small-angle X-ray scattering profiles, it is possible to assess within seconds whether a cell culture contains microcrystals, even in a small percentage of cells. Since such information cannot be obtained by other established detection methods in this time frame, this screening approach has the potential to overcome one of the bottlenecks of intracellular crystal detection. Moreover, the association of the Bragg peak positions in the scattering curves with the unit-cell composition of the protein crystals raises the possibility of investigating the impact of environmental conditions on the crystal structure of the intracellular protein crystals. This information provides valuable insights helping to further understand the in cellulo crystallization process.

show abstract

Section: Introductionmentioning

confidence: 99%

Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Очевидно, что все модели, предусматривающие образование структурно-оформленных (полиэдрических) кластеров-предшественников, требуют весьма тщательного изучения разными методами процессов их образования и существования в кристаллообразующих средах. Интересно, что недавно были опубликованы работы (Kovalchuk et al, 2016;Дьякова и др., 2017), в которых показано, что кластеры-прекурсоры представляют собой 3D-фрагменты макро кристаллической структуры. Эти данные были получены с помощью малоуглового рассеяния рентгеновских лучей растворами.…”

Section: предзародышевые кластеры и неклассические модели зародышеобрunclassified

Pre-nucleation clusters and non-classical crystal formation

Askhabov

Магомедович²

2019

Zapiski RMO

View full text Add to dashboard Cite

“…1). The procedure for model selection is described in Kovalchuk et al (2016). The asymmetric unit was reproduced by the symmetry operators using Coot (Emsley et al, 2010).…”

Section: Sans Experiments and Data Processingmentioning

confidence: 99%

“…For experimental data processing, we used oligomeric models of lysozyme derived from the crystal structure and our previous work (Kovalchuk et al, 2016;Marchenkova et al, 2016). Using these models, it is possible to determine the volume fractions of oligomers in the protein solutions under different conditions.…”

Section: Introductionmentioning

confidence: 99%

Octamer formation in lysozyme solutions at the initial crystallization stage detected by small-angle neutron scattering

Бойкова

Dyakova

Ilina

et al. 2017

Acta Crystallogr D Struct Biol

Self Cite

View full text Add to dashboard Cite

Solutions of lysozyme in heavy water were studied by small-angle neutron scattering (SANS) at concentrations of 40, 20 and 10 mg ml with and without the addition of precipitant, and at temperatures of 10, 20 and 30°C. In addition to the expected protein monomers, dimeric and octameric species were identified in solutions at the maximum concentration and close to the optimal conditions for crystallization. An optimal temperature for octamer formation was identified and both deviation from this temperature and a reduction in protein concentration led to a significant decrease in the volume fractions of octamers detected. In the absence of precipitant, only monomers and a minor fraction of dimers are present in solution.

show abstract

Investigation of the Initial Crystallization Stage in Lysozyme Solutions by Small-Angle X-ray Scattering

Cited by 56 publications

References 33 publications

Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

Pre-nucleation clusters and non-classical crystal formation

Octamer formation in lysozyme solutions at the initial crystallization stage detected by small-angle neutron scattering

Contact Info

Product

Resources

About