Cargando…

Single-step purification of full-length human androgen receptor

The full-length human androgen receptor with an N-terminal biotin acceptor peptide tag was overexpressed in Spodoptera frugiperda cells in the presence of 1 µM dihydrotestosterone. Site-specific biotinylation of BAP was achieved in vivo by co-expression of E. coli biotin holoenzyme synthetase. The a...

Descripción completa

Detalles Bibliográficos
Autores principales: Juzumiene, Dalia, Chang, Ching-yi, Fan, Daju, Hartney, Tanya, Norris, John D., McDonnell, Donald P.
Formato: Texto
Lenguaje:English
Publicado: The Nuclear Receptor Signaling Atlas 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1402216/
https://www.ncbi.nlm.nih.gov/pubmed/16604169
http://dx.doi.org/10.1621/nrs.03001
_version_ 1782126997923692544
author Juzumiene, Dalia
Chang, Ching-yi
Fan, Daju
Hartney, Tanya
Norris, John D.
McDonnell, Donald P.
author_facet Juzumiene, Dalia
Chang, Ching-yi
Fan, Daju
Hartney, Tanya
Norris, John D.
McDonnell, Donald P.
author_sort Juzumiene, Dalia
collection PubMed
description The full-length human androgen receptor with an N-terminal biotin acceptor peptide tag was overexpressed in Spodoptera frugiperda cells in the presence of 1 µM dihydrotestosterone. Site-specific biotinylation of BAP was achieved in vivo by co-expression of E. coli biotin holoenzyme synthetase. The androgen receptor was purified by single-step affinity chromatography using Streptavidin Mutein Matrix under native conditions. The resultant protein was active, stable, 95% homogeneous, and we obtained sufficient yield for use in functional and structural studies.
format Text
id pubmed-1402216
institution National Center for Biotechnology Information
language English
publishDate 2005
publisher The Nuclear Receptor Signaling Atlas
record_format MEDLINE/PubMed
spelling pubmed-14022162006-04-06 Single-step purification of full-length human androgen receptor Juzumiene, Dalia Chang, Ching-yi Fan, Daju Hartney, Tanya Norris, John D. McDonnell, Donald P. Nucl Recept Signal Methods The full-length human androgen receptor with an N-terminal biotin acceptor peptide tag was overexpressed in Spodoptera frugiperda cells in the presence of 1 µM dihydrotestosterone. Site-specific biotinylation of BAP was achieved in vivo by co-expression of E. coli biotin holoenzyme synthetase. The androgen receptor was purified by single-step affinity chromatography using Streptavidin Mutein Matrix under native conditions. The resultant protein was active, stable, 95% homogeneous, and we obtained sufficient yield for use in functional and structural studies. The Nuclear Receptor Signaling Atlas 2005-10-21 /pmc/articles/PMC1402216/ /pubmed/16604169 http://dx.doi.org/10.1621/nrs.03001 Text en Copyright © 2005, Juzumiene et al. This is an open-access article distributed under the terms of the Creative Commons Non-Commercial Attribution License, which permits unrestricted non-commercial use distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods
Juzumiene, Dalia
Chang, Ching-yi
Fan, Daju
Hartney, Tanya
Norris, John D.
McDonnell, Donald P.
Single-step purification of full-length human androgen receptor
title Single-step purification of full-length human androgen receptor
title_full Single-step purification of full-length human androgen receptor
title_fullStr Single-step purification of full-length human androgen receptor
title_full_unstemmed Single-step purification of full-length human androgen receptor
title_short Single-step purification of full-length human androgen receptor
title_sort single-step purification of full-length human androgen receptor
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1402216/
https://www.ncbi.nlm.nih.gov/pubmed/16604169
http://dx.doi.org/10.1621/nrs.03001
work_keys_str_mv AT juzumienedalia singlesteppurificationoffulllengthhumanandrogenreceptor
AT changchingyi singlesteppurificationoffulllengthhumanandrogenreceptor
AT fandaju singlesteppurificationoffulllengthhumanandrogenreceptor
AT hartneytanya singlesteppurificationoffulllengthhumanandrogenreceptor
AT norrisjohnd singlesteppurificationoffulllengthhumanandrogenreceptor
AT mcdonnelldonaldp singlesteppurificationoffulllengthhumanandrogenreceptor