Positive and negative effects of graphite flake and monolayer graphene oxide templates on protein crystallization (Open Access)--颗粒学报PARTICUOLOGY

Lin, Z., Tian, W., Su, Y., Yadav, V. K., & Yang, H. (2023). Positive and negative effects of graphite flake and monolayer graphene oxide templates on protein crystallization. Particuology, 79, 27-34. https://doi.org/10.1016/j.partic.2022.10.014

Positive and negative effects of graphite flake and monolayer graphene oxide templates on protein crystallization (Open Access)

Zhichun Lin ^a, Wenqing Tian ^a, Yang Su ^b, Vikash Kumar Yadav ^c, Huaiyu Yang ^a *

a Department of Chemical Engineering, Loughborough University, Loughborough, LE11 3TU, UK
b Institute of Materials Research, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, 518055, China
c Human Sciences Research Centre, University of Derby, DE22 1GB, UK

10.1016/j.partic.2022.10.014

Volume 79, August 2023, Pages 27-34

Received 12 August 2022, Revised 10 October 2022, Accepted 27 October 2022, Available online 11 November 2022, Version of Record 24 January 2023.

E-mail: H.yang3@lboro.ac.uk

Highlights

• Positive or negative effects of nano monolayer graphite oxide templates on protein crystallization.

• Shorter nucleation times with graphite templates, and preference of nucleation on the edge of the templates.

• Graphite oxide templates enfolded inside the protein crystals.

• Crystal numbers in the droplets: with graphite template > with graphene oxide templates > without templates.

• Average crystal sizes in the droplets as opposite order as crystal numbers.

Abstract

Heterogeneous template-induced nucleation is a promising way to regulate protein crystallization events and could be employed for purification processes and crystallographic studies. Protein crystallization process with graphite and graphene oxide, as heterogeneous templates, were investigated. More than 640 hanging drops with different concentrations of Lysozyme (30, 50, 70, 100 mg/mL) and NaCl (0.7, 0.9, 1.1, 1.3, 1.5 M) were crystallised at 4 °C with or without graphite/graphene oxide templates. The induction times and crystallization process were observed under the microscope. The lysozyme in the solutions with graphite flakes nucleated faster under all the conditions than the lysozyme with equal experimental conditions without templates. The crystals preferred to grow around the edge of graphite flakes than on the flat surfaces. In the droplets with monolayer graphene oxide, more crystals appeared around graphene oxide particles, and the faster or slower nucleation processes with templates were dependent on the lysozyme and NaCl concentrations. Graphene oxide templates strongly inhibited nucleation at high lysozyme concentrations but promoted nucleation at low lysozyme concentrations. Both heterogeneous templates changed the crystal morphology and the crystallization kinetics. More crystals were observed in the solution with graphite templatesthan with graphene oxide templates and without any template.

Graphical abstract

Keywords

Graphite; Graphene oxide; Protein crystallization; Adsorption; Heterogeneous nucleation; Template technology

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