Cargando…
Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization
Structural biology and structural genomics projects routinely rely on recombinantly expressed proteins, but many proteins and complexes are difficult to obtain by this approach. We investigated native source proteins for high-throughput protein crystallography applications. The Escherichia coli prot...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3290569/ https://www.ncbi.nlm.nih.gov/pubmed/22393408 http://dx.doi.org/10.1371/journal.pone.0032498 |
_version_ | 1782225018573291520 |
---|---|
author | Totir, Monica Echols, Nathaniel Nanao, Max Gee, Christine L. Moskaleva, Alisa Gradia, Scott Iavarone, Anthony T. Berger, James M. May, Andrew P. Zubieta, Chloe Alber, Tom |
author_facet | Totir, Monica Echols, Nathaniel Nanao, Max Gee, Christine L. Moskaleva, Alisa Gradia, Scott Iavarone, Anthony T. Berger, James M. May, Andrew P. Zubieta, Chloe Alber, Tom |
author_sort | Totir, Monica |
collection | PubMed |
description | Structural biology and structural genomics projects routinely rely on recombinantly expressed proteins, but many proteins and complexes are difficult to obtain by this approach. We investigated native source proteins for high-throughput protein crystallography applications. The Escherichia coli proteome was fractionated, purified, crystallized, and structurally characterized. Macro-scale fermentation and fractionation were used to subdivide the soluble proteome into 408 unique fractions of which 295 fractions yielded crystals in microfluidic crystallization chips. Of the 295 crystals, 152 were selected for optimization, diffraction screening, and data collection. Twenty-three structures were determined, four of which were novel. This study demonstrates the utility of native source proteins for high-throughput crystallography. |
format | Online Article Text |
id | pubmed-3290569 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-32905692012-03-05 Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization Totir, Monica Echols, Nathaniel Nanao, Max Gee, Christine L. Moskaleva, Alisa Gradia, Scott Iavarone, Anthony T. Berger, James M. May, Andrew P. Zubieta, Chloe Alber, Tom PLoS One Research Article Structural biology and structural genomics projects routinely rely on recombinantly expressed proteins, but many proteins and complexes are difficult to obtain by this approach. We investigated native source proteins for high-throughput protein crystallography applications. The Escherichia coli proteome was fractionated, purified, crystallized, and structurally characterized. Macro-scale fermentation and fractionation were used to subdivide the soluble proteome into 408 unique fractions of which 295 fractions yielded crystals in microfluidic crystallization chips. Of the 295 crystals, 152 were selected for optimization, diffraction screening, and data collection. Twenty-three structures were determined, four of which were novel. This study demonstrates the utility of native source proteins for high-throughput crystallography. Public Library of Science 2012-02-29 /pmc/articles/PMC3290569/ /pubmed/22393408 http://dx.doi.org/10.1371/journal.pone.0032498 Text en Totir 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 Totir, Monica Echols, Nathaniel Nanao, Max Gee, Christine L. Moskaleva, Alisa Gradia, Scott Iavarone, Anthony T. Berger, James M. May, Andrew P. Zubieta, Chloe Alber, Tom Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title | Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title_full | Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title_fullStr | Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title_full_unstemmed | Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title_short | Macro-to-Micro Structural Proteomics: Native Source Proteins for High-Throughput Crystallization |
title_sort | macro-to-micro structural proteomics: native source proteins for high-throughput crystallization |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3290569/ https://www.ncbi.nlm.nih.gov/pubmed/22393408 http://dx.doi.org/10.1371/journal.pone.0032498 |
work_keys_str_mv | AT totirmonica macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT echolsnathaniel macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT nanaomax macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT geechristinel macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT moskalevaalisa macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT gradiascott macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT iavaroneanthonyt macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT bergerjamesm macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT mayandrewp macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT zubietachloe macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization AT albertom macrotomicrostructuralproteomicsnativesourceproteinsforhighthroughputcrystallization |