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Improved success of sparse matrix protein crystallization screening with heterogeneous nucleating agents

Journal Article


Abstract


  • 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. © 2007 Thakur et al.

UOW Authors


  •   Thakur, Anil (external author)
  •   Robin, Gautier (external author)
  •   Guncar, Gregor (external author)
  •   Saunders, Neil (external author)
  •   Newman, Janet (external author)
  •   Martin, Jennifer (external author)
  •   Kobe, Bostjan (external author)

Publication Date


  • 2007

Citation


  • Thakur, A. S., Robin, G., Guncar, G., Saunders, N. F. W., Newman, J., Martin, J. L. & Kobe, B. (2007). Improved success of sparse matrix protein crystallization screening with heterogeneous nucleating agents. PLoS One, 2 (10), e1091-1-e1091-6.

Scopus Eid


  • 2-s2.0-42449125742

Start Page


  • e1091-1

End Page


  • e1091-6

Volume


  • 2

Issue


  • 10

Place Of Publication


  • United States

Abstract


  • 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. © 2007 Thakur et al.

UOW Authors


  •   Thakur, Anil (external author)
  •   Robin, Gautier (external author)
  •   Guncar, Gregor (external author)
  •   Saunders, Neil (external author)
  •   Newman, Janet (external author)
  •   Martin, Jennifer (external author)
  •   Kobe, Bostjan (external author)

Publication Date


  • 2007

Citation


  • Thakur, A. S., Robin, G., Guncar, G., Saunders, N. F. W., Newman, J., Martin, J. L. & Kobe, B. (2007). Improved success of sparse matrix protein crystallization screening with heterogeneous nucleating agents. PLoS One, 2 (10), e1091-1-e1091-6.

Scopus Eid


  • 2-s2.0-42449125742

Start Page


  • e1091-1

End Page


  • e1091-6

Volume


  • 2

Issue


  • 10

Place Of Publication


  • United States