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Updated: Jul 10, 2026
Improving the Success Rate of Protein Crystallization by Random Microseed Matrix Screening
Published on: August 31, 2013
Improved success of sparse matrix protein crystallization screening with heterogeneous nucleating agents
Anil S Thakur1, Gautier Robin, Gregor Guncar
1School of Molecular and Microbial Sciences, University of Queensland, St. Lucia, Brisbane, Queensland, Australia.
Background:
Crystallization is a major bottleneck in the process of macromolecular structure determination by X-ray crystallography. Successful crystallization requires the formation of nuclei and their subsequent growth to crystals of suitable size. Crystal growth generally occurs spontaneously in a supersaturated solution as a result of homogenous nucleation. However, in a typical sparse matrix screening experiment, precipitant and protein concentration are not sampled extensively, and supersaturation conditions suitable for nucleation are often missed.
Methodology/Principal Findings:
We tested the effect of nine potential heterogenous nucleating agents on crystallization of ten test proteins in a sparse matrix screen. Several nucleating agents induced crystal formation under conditions where no crystallization occurred in the absence of the nucleating agent. Four nucleating agents: dried seaweed; horse hair; cellulose and hydroxyapatite, had a considerable overall positive effect on crystallization success. This effect was further enhanced when these nucleating agents were used in combination with each other.
Conclusions/Significance:
Our results suggest that the addition of heterogeneous nucleating agents increases the chances of crystal formation when using sparse matrix screens.
