Cargando…

Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Thakur, Anil S., Robin, Gautier, Guncar, Gregor, Saunders, Neil F. W., Newman, Janet, Martin, Jennifer L., Kobe, Bostjan
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2034409/
https://www.ncbi.nlm.nih.gov/pubmed/17971854
http://dx.doi.org/10.1371/journal.pone.0001091
_version_ 1782137004894453760
author Thakur, Anil S.
Robin, Gautier
Guncar, Gregor
Saunders, Neil F. W.
Newman, Janet
Martin, Jennifer L.
Kobe, Bostjan
author_facet Thakur, Anil S.
Robin, Gautier
Guncar, Gregor
Saunders, Neil F. W.
Newman, Janet
Martin, Jennifer L.
Kobe, Bostjan
author_sort Thakur, Anil S.
collection PubMed
description 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.
format Text
id pubmed-2034409
institution National Center for Biotechnology Information
language English
publishDate 2007
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-20344092007-10-31 Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents Thakur, Anil S. Robin, Gautier Guncar, Gregor Saunders, Neil F. W. Newman, Janet Martin, Jennifer L. Kobe, Bostjan PLoS One Research Article 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. Public Library of Science 2007-10-31 /pmc/articles/PMC2034409/ /pubmed/17971854 http://dx.doi.org/10.1371/journal.pone.0001091 Text en Thakur et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Thakur, Anil S.
Robin, Gautier
Guncar, Gregor
Saunders, Neil F. W.
Newman, Janet
Martin, Jennifer L.
Kobe, Bostjan
Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title_full Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title_fullStr Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title_full_unstemmed Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title_short Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents
title_sort improved success of sparse matrix protein crystallization screening with heterogeneous nucleating agents
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2034409/
https://www.ncbi.nlm.nih.gov/pubmed/17971854
http://dx.doi.org/10.1371/journal.pone.0001091
work_keys_str_mv AT thakuranils improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT robingautier improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT guncargregor improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT saundersneilfw improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT newmanjanet improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT martinjenniferl improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents
AT kobebostjan improvedsuccessofsparsematrixproteincrystallizationscreeningwithheterogeneousnucleatingagents