Rapid and efficient purification of Drosophila homeodomain transcription factors for biophysical characterization.

Animals Drosophila Proteins / chemistry Drosophila melanogaster Homeodomain Proteins / chemistry Recombinant Fusion Proteins / chemistry
DNA binding Electrophoretic mobility shift assay Homeodomain transcription factor NMR spectroscopy Protein-DNA interactions

Journal

Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496

Informations de publication

Date de publication:
06 2019
Historique:
received: 04 01 2019
accepted: 03 02 2019
pubmed: 11 2 2019
medline: 9 4 2020
entrez: 11 2 2019
Statut: ppublish

Résumé

Homeodomain transcription factors (HD TFs) are a large class of evolutionarily conserved DNA binding proteins that contain a basic 60-amino acid region required for binding to specific DNA sites. In Drosophila melanogaster, many of these HD TFs are expressed in the early embryo and control transcription of target genes in development through their interaction with cis-regulatory modules. Previous studies where some of the Drosophila HD TFs were purified required the use of strong denaturants (i.e. 6 M urea) and multiple chromatography columns, making the downstream biochemical examination of the isolated protein difficult. To circumvent these obstacles, we have developed a streamlined expression and purification protocol to produce large yields of Drosophila HD TFs. Using the HD TFs FUSHI-TARAZU (FTZ), ANTENNAPEDIA (ANTP), ABDOMINAL-A (ABD-A), ABDOMINAL-B (ABD-B), and ULTRABITHORAX (UBX) as examples, we demonstrate that our 3-day protocol involving the overexpression of His

Identifiants

DOI: 10.1016/j.pep.2019.02.001 PMID: 30738927 PMC: PMC6424608
pubmed: 30738927
pii: S1046-5928(19)30006-3
doi: 10.1016/j.pep.2019.02.001
pmc: PMC6424608
mid: NIHMS1521620
pii:
doi:

Substances chimiques

Drosophila Proteins 0
Homeodomain Proteins 0
Recombinant Fusion Proteins 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

9-14

Subventions

Organisme : NIGMS NIH HHS
ID : R15 GM110571
Pays : United States
Organisme : NIGMS NIH HHS
ID : R15 GM126432
Pays : United States

Informations de copyright

Copyright © 2019 Elsevier Inc. All rights reserved.

Références

Cell. 1990 Nov 2;63(3):579-90
pubmed: 1977522
Proc Natl Acad Sci U S A. 2005 Apr 5;102(14):4936-42
pubmed: 15788537
Adv Exp Med Biol. 2010;689:17-40
pubmed: 20795320
Proc Natl Acad Sci U S A. 1990 Jun;87(11):4093-7
pubmed: 1971945
J Biomol NMR. 1995 Nov;6(3):277-93
pubmed: 8520220
Nature. 1997 Apr 10;386(6625):569-77
pubmed: 9121580
Mol Cell Biol. 1988 Nov;8(11):4598-607
pubmed: 2905420
Chromosoma. 2016 Jun;125(3):497-521
pubmed: 26464018
J Mol Biol. 1994 May 6;238(3):333-45
pubmed: 7909851
EMBO J. 1992 Mar;11(3):991-1002
pubmed: 1347746
Nucleic Acids Res. 2001 Aug 1;29(15):3258-69
pubmed: 11470884
Mech Dev. 2001 Sep;107(1-2):39-53
pubmed: 11520662
Nat Biotechnol. 2005 Aug;23(8):988-94
pubmed: 16041365
Nature. 1997 Feb 6;385(6616):552-5
pubmed: 9020364
Pediatr Res. 1997 Oct;42(4):421-9
pubmed: 9380431
EMBO J. 1988 Dec 20;7(13):4305-9
pubmed: 2907480
Cell. 1988 Sep 23;54(7):1081-90
pubmed: 3046753
Cell. 2008 Jun 27;133(7):1277-89
pubmed: 18585360
Development. 1997 May;124(10):1851-64
pubmed: 9169833
Bioinformatics. 2007 Nov 1;23(21):2947-8
pubmed: 17846036
Hum Genet. 2014 Jun;133(6):701-11
pubmed: 24390306
J Biomol NMR. 1994 Sep;4(5):603-14
pubmed: 22911360
Cell. 1989 Nov 3;59(3):563-71
pubmed: 2572328
Mol Cell Biol. 1991 Jul;11(7):3613-23
pubmed: 1675428
Nature. 2001 Feb 15;409(6822):832-3
pubmed: 11237001
J Mol Biol. 1993 Dec 20;234(4):1084-93
pubmed: 7903398
Curr Opin Genet Dev. 2017 Apr;43:1-8
pubmed: 27768937
PLoS Genet. 2009 Nov;5(11):e1000709
pubmed: 19893611
J Mol Biol. 1990 Jul 5;214(1):183-97
pubmed: 2164583
EMBO J. 1988 Dec 20;7(13):4299-304
pubmed: 2907479
Dev Biol. 2011 Nov 15;359(2):290-302
pubmed: 21821017
Cell. 2004 Jan 23;116(2):247-57
pubmed: 14744435
Mech Dev. 2014 Feb;131:68-77
pubmed: 24514265
Nucleic Acids Res. 2008 May;36(8):2547-60
pubmed: 18332042
Bioinformatics. 2009 May 1;25(9):1189-91
pubmed: 19151095
J Biomol NMR. 2008 May;41(1):5-7
pubmed: 18398685
Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10738-42
pubmed: 1359544
Nature. 1984 Mar 29-Apr 4;308(5958):428-33
pubmed: 6323992
EMBO J. 1990 Dec;9(12):3967-74
pubmed: 1979032
Development. 2003 Dec;130(26):6569-75
pubmed: 14660545
New Biol. 1990 Feb;2(2):171-8
pubmed: 1982071
EMBO J. 1990 Oct;9(10):3085-92
pubmed: 1976507
PLoS Biol. 2008 Feb;6(2):e27
pubmed: 18271625
Nat Protoc. 2007;2(8):1849-61
pubmed: 17703195
Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1450-4
pubmed: 8434005
Curr Top Dev Biol. 2012;98:97-120
pubmed: 22305160
Development. 2010 Jan;137(1):5-13
pubmed: 20023155
Nucleic Acids Res. 2009 Jul;37(Web Server issue):W202-8
pubmed: 19458158
Cell Rep. 2018 Aug 28;24(9):2221-2230
pubmed: 30157419

Auteurs

Rachel Orlomoski (R)

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA; Department of Biology, Clark University, 950 Main St, Worcester, MA, 01610, USA.

Aaron Bogle (A)

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA; Department of Biology, Clark University, 950 Main St, Worcester, MA, 01610, USA.

Jeanmarie Loss (J)

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA; Department of Biology, Clark University, 950 Main St, Worcester, MA, 01610, USA.

Rylee Simons (R)

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA; Department of Biology, Clark University, 950 Main St, Worcester, MA, 01610, USA.

Jacqueline M Dresch (JM)

Department of Math & Computer Science, Clark University, 950 Main St, Worcester, MA, 01610, USA.

Robert A Drewell (RA)

Department of Biology, Clark University, 950 Main St, Worcester, MA, 01610, USA. Electronic address: rdrewell@clarku.edu.

Donald E Spratt (DE)

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA. Electronic address: dspratt@clarku.edu.

Articles similaires

Evaluating the efficacy of telesurgery with dual console SSI Mantra Surgical Robotic System: experiment on animal model and clinical trials.

Sudhir Prem Srivastava, Vishwajyoti Pascual Srivastava, Avinesh Singh et al.
1.00
Robotic Surgical Procedures Animals Humans Telemedicine Models, Animal

Odour generalisation and detection dog training.

Lyn Caldicott, Thomas W Pike, Helen E Zulch et al.
1.00
Animals Odorants Dogs Generalization, Psychological Smell

FBXO22 inhibits colitis and colorectal carcinogenesis by regulating the degradation of the S2448-phosphorylated form of mTOR.

Minle Li, Xuan Chen, Pengfei Qu et al.
1.00
Animals TOR Serine-Threonine Kinases Colorectal Neoplasms Colitis Mice

Use of organic material provided by an automatic enrichment device by weaner pigs and its influence on tail lesions.

Philipp Heseker, Jeanette Probst, Stefanie Ammer et al.
1.00
Animals Tail Swine Behavior, Animal Animal Husbandry

Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Rapid and efficient purification of Drosophila...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines d'arthropode › Protéines d'insecte › Protéines de Drosophila › Rapid and efficient purification of Drosophila...
questionsmedicales.fr › Eucaryotes › Animaux › Invertébrés › Arthropodes › Insectes › Pterygota › Neoptera › Holometabola › Diptera › Drosophilidae › Drosophila › Drosophila melanogaster › Rapid and efficient purification of Drosophila...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de liaison à l'ADN › Protéines à homéodomaine › Rapid and efficient purification of Drosophila...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines recombinantes › Protéines de fusion recombinantes › Rapid and efficient purification of Drosophila...