Enhancement of nucleation during hanging drop protein crystallization using HF Treatment of cover glasses

Guo, Yang; Yin, Da‐Chuan; Lu, Qi; Wang, Xi‐Kai; Li, Jun

doi:10.1002/crat.200900609

Cited by 10 publications

(13 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compared with a smooth surface, the surfaces of the oxidized plates and the control plates both had much higher surface areas and many more small nanopores. Theoretically, the free-energy barrier for nucleation is decreased at a higher roughness (Guo et al, 2010;Liu et al, 2007). As shown in Table 3, the surface of the oxidized plates was smoother than that of the untreated plates.…”

Section: Larger Surface Area Could Promote the Evaporationmentioning

confidence: 85%

See 1 more Smart Citation

Surface treatment by oxidizing the plates can alter the response of protein crystallization

et al. 2014

Self Cite

View full text Add to dashboard Cite

This report describes the modification of crystallization plates by simply oxidizing the surface of the protein wells. The oxidized crystallization plates were tested in standard protein crystallization screening and reproducibility studies. The results showed that the protein wells of the treated plates were smoother and more optically transparent than those of the untreated plates, and more importantly, protein crystallization was significantly promoted after the oxidation treatment. Because there is no change to the routine screening protocol, this method is simple and easy to apply in protein crystallization.

show abstract

Section: Larger Surface Area Could Promote the Evaporationmentioning

confidence: 85%

“…5, we could compare the contact areas between the OCP and control groups. The contact area was larger for the OCP than the control plates, and the larger contact area provided more opportunities for nucleation (Guo et al, 2010). Thus, there would be a greater likelihood of obtaining crystals in the OCP group.…”

Section: Discussionmentioning

confidence: 97%

Surface treatment by oxidizing the plates can alter the response of protein crystallization

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…Schmidt (8) argued that hanging drop or sitting drop vapor diffusion is a reasonably good screening method for identifying growth potential of a protein crystal. Since the protein crystallization process involves consecutive stages of nucleation and growth, successful crystallization first required nucleation (9). To this end, the hanging drop method serves as a robust technique for forming protein crystals.…”

Section: Introductionmentioning

confidence: 99%

Verification of McCabe's ΔL Law for Growth of Lysozyme Crystals by Hanging Drop Method

Sengupta

Farrell

2012

Separation Science and Technology

View full text Add to dashboard Cite

Lysozyme is a naturally occurring enzyme in egg white and has high commercial importance due to its antimicrobial properties. The main objective of this work was to study the growth rate of lysozyme crystals isolated from egg for the first 72 hours and verify the results with McCabe's constant crystal growth theory. The separation of lysozyme was carried out by adsorption onto a cationic Amberlite resin using phosphate buffer at pH 4.8. Hanging drop crystallization method was used to form high purity lysozyme crystals from the embryonic stage. To this end, this work differs from an earlier work of Forsythe et al., who used seed crystals in the size range of 10 lm-40 lm for face growth measurements at different pH values. The maximum crystal size recorded in the present work was 392.86 lm, which is within the typical size range of 50 lm-500 lm for which constant crystal growth is expected to hold according to McCabe's DL law. Electron micrographs (SEM) revealed the structure and dimensions of the crystals while SDS-Page was used to measure the purity of the crystals. The SEM results showed that lysozyme growth rate was linear and agreed with McCabe's constant growth theory, producing a growth rate of 1.77 Â 10 À3 mm Á s À1 .

show abstract

“…Mineral substrate [11], silanised mica surface [12], lipid bilayers deposited on glass cover slips (for membrane protein crystallisation) [13,14], polymeric film containing poly- l -lysine or poly- l -aspartate ionisable groups [15], and a modified surface with a varying roughness [16,17] have all been proven to be useful for promoting protein crystallisation. These findings demonstrated that heterogeneous nucleation can effectively occur on large surfaces.…”

Section: Introductionmentioning

confidence: 99%

An Investigation of the Effects of Self-Assembled Monolayers on Protein Crystallisation

Zhang

Shen

Wang

et al. 2013

IJMS

Self Cite

View full text Add to dashboard Cite

Most protein crystallisation begins from heterogeneous nucleation; in practice, crystallisation typically occurs in the presence of a solid surface in the solution. The solid surface provides a nucleation site such that the energy barrier for nucleation is lower on the surface than in the bulk solution. Different types of solid surfaces exhibit different surface energies, and the nucleation barriers depend on the characteristics of the solid surfaces. Therefore, treatment of the solid surface may alter the surface properties to increase the chance to obtain protein crystals. In this paper, we propose a method to modify the glass cover slip using a self-assembled monolayer (SAM) of functional groups (methyl, sulfydryl and amino), and we investigated the effect of each SAM on protein crystallisation. The results indicated that both crystallisation success rate in a reproducibility study, and crystallisation hits in a crystallisation screening study, were increased using the SAMs, among which, the methyl-modified SAM demonstrated the most significant improvement. These results illustrated that directly modifying the crystallisation plates or glass cover slips to create surfaces that favour heterogeneous nucleation can be potentially useful in practical protein crystallisation, and the utilisation of a SAM containing a functional group can be considered a promising technique for the treatment of the surfaces that will directly contact the crystallisation solution.

show abstract

Enhancement of nucleation during hanging drop protein crystallization using HF Treatment of cover glasses

Cited by 10 publications

References 22 publications

Surface treatment by oxidizing the plates can alter the response of protein crystallization

Surface treatment by oxidizing the plates can alter the response of protein crystallization

Verification of McCabe's ΔL Law for Growth of Lysozyme Crystals by Hanging Drop Method

An Investigation of the Effects of Self-Assembled Monolayers on Protein Crystallisation

Contact Info

Product

Resources

About