Purification, crystallization and preliminary X-ray diffraction analysis of the HMG domain of Sox17 in complex with DNA

Calista Keow Leng Ng, Paaventhan Palasingam, Rajakannan Venkatachalam, Nithya Baburajendran, Jason Cheng, Ralf Jauch, Prasanna Kolatkar

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Sox17 is a member of the SRY-related high-mobility group (HMG) of transcription factors that have been shown to direct endodermal differentiation in early mammalian development. The LAMA1 gene encoding the α-chain of laminin-1 has been reported to be directly bound and regulated by Sox17. This paper describes the details of initial crystallization attempts with the HMG domain of mouse Sox17 (mSox17-HMG) with a 16-mer DNA element derived from the LAMA1 enhancer and optimization strategies to obtain a better diffracting crystal. The best diffracting crystal was obtained in a condition containing 0.1 M Tris-HCl pH 7.4, 0.2 M MgCl2, 30% PEG 3350 using the hanging-drop vapour-diffusion method. A highly redundant in-house data set was collected to 2.75 Å resolution with 99% completeness. The presence of the mSox17-HMG-DNA complex within the crystals was confirmed and Matthews analysis indicated the presence of one complex per asymmetric unit.

Original languageEnglish
Pages (from-to)1184-1187
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume64
Issue number12
DOIs
Publication statusPublished - 28 Nov 2008
Externally publishedYes

Fingerprint

Crystallization
purification
X-Ray Diffraction
X ray diffraction analysis
Purification
deoxyribonucleic acid
crystallization
Crystals
Magnesium Chloride
DNA
diffraction
Gene encoding
x rays
Transcription Factors
crystals
completeness
Vapors
genes
Genes
mice

Keywords

  • HMG domains
  • LAMA1
  • Sox17

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

Purification, crystallization and preliminary X-ray diffraction analysis of the HMG domain of Sox17 in complex with DNA. / Ng, Calista Keow Leng; Palasingam, Paaventhan; Venkatachalam, Rajakannan; Baburajendran, Nithya; Cheng, Jason; Jauch, Ralf; Kolatkar, Prasanna.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 64, No. 12, 28.11.2008, p. 1184-1187.

Research output: Contribution to journalArticle

Ng, Calista Keow Leng ; Palasingam, Paaventhan ; Venkatachalam, Rajakannan ; Baburajendran, Nithya ; Cheng, Jason ; Jauch, Ralf ; Kolatkar, Prasanna. / Purification, crystallization and preliminary X-ray diffraction analysis of the HMG domain of Sox17 in complex with DNA. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2008 ; Vol. 64, No. 12. pp. 1184-1187.
@article{4b85e742b4e4408e89738ba88034850c,
title = "Purification, crystallization and preliminary X-ray diffraction analysis of the HMG domain of Sox17 in complex with DNA",
abstract = "Sox17 is a member of the SRY-related high-mobility group (HMG) of transcription factors that have been shown to direct endodermal differentiation in early mammalian development. The LAMA1 gene encoding the α-chain of laminin-1 has been reported to be directly bound and regulated by Sox17. This paper describes the details of initial crystallization attempts with the HMG domain of mouse Sox17 (mSox17-HMG) with a 16-mer DNA element derived from the LAMA1 enhancer and optimization strategies to obtain a better diffracting crystal. The best diffracting crystal was obtained in a condition containing 0.1 M Tris-HCl pH 7.4, 0.2 M MgCl2, 30{\%} PEG 3350 using the hanging-drop vapour-diffusion method. A highly redundant in-house data set was collected to 2.75 {\AA} resolution with 99{\%} completeness. The presence of the mSox17-HMG-DNA complex within the crystals was confirmed and Matthews analysis indicated the presence of one complex per asymmetric unit.",
keywords = "HMG domains, LAMA1, Sox17",
author = "Ng, {Calista Keow Leng} and Paaventhan Palasingam and Rajakannan Venkatachalam and Nithya Baburajendran and Jason Cheng and Ralf Jauch and Prasanna Kolatkar",
year = "2008",
month = "11",
day = "28",
doi = "10.1107/S1744309108038724",
language = "English",
volume = "64",
pages = "1184--1187",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",
number = "12",

}

TY - JOUR

T1 - Purification, crystallization and preliminary X-ray diffraction analysis of the HMG domain of Sox17 in complex with DNA

AU - Ng, Calista Keow Leng

AU - Palasingam, Paaventhan

AU - Venkatachalam, Rajakannan

AU - Baburajendran, Nithya

AU - Cheng, Jason

AU - Jauch, Ralf

AU - Kolatkar, Prasanna

PY - 2008/11/28

Y1 - 2008/11/28

N2 - Sox17 is a member of the SRY-related high-mobility group (HMG) of transcription factors that have been shown to direct endodermal differentiation in early mammalian development. The LAMA1 gene encoding the α-chain of laminin-1 has been reported to be directly bound and regulated by Sox17. This paper describes the details of initial crystallization attempts with the HMG domain of mouse Sox17 (mSox17-HMG) with a 16-mer DNA element derived from the LAMA1 enhancer and optimization strategies to obtain a better diffracting crystal. The best diffracting crystal was obtained in a condition containing 0.1 M Tris-HCl pH 7.4, 0.2 M MgCl2, 30% PEG 3350 using the hanging-drop vapour-diffusion method. A highly redundant in-house data set was collected to 2.75 Å resolution with 99% completeness. The presence of the mSox17-HMG-DNA complex within the crystals was confirmed and Matthews analysis indicated the presence of one complex per asymmetric unit.

AB - Sox17 is a member of the SRY-related high-mobility group (HMG) of transcription factors that have been shown to direct endodermal differentiation in early mammalian development. The LAMA1 gene encoding the α-chain of laminin-1 has been reported to be directly bound and regulated by Sox17. This paper describes the details of initial crystallization attempts with the HMG domain of mouse Sox17 (mSox17-HMG) with a 16-mer DNA element derived from the LAMA1 enhancer and optimization strategies to obtain a better diffracting crystal. The best diffracting crystal was obtained in a condition containing 0.1 M Tris-HCl pH 7.4, 0.2 M MgCl2, 30% PEG 3350 using the hanging-drop vapour-diffusion method. A highly redundant in-house data set was collected to 2.75 Å resolution with 99% completeness. The presence of the mSox17-HMG-DNA complex within the crystals was confirmed and Matthews analysis indicated the presence of one complex per asymmetric unit.

KW - HMG domains

KW - LAMA1

KW - Sox17

UR - http://www.scopus.com/inward/record.url?scp=57349105289&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57349105289&partnerID=8YFLogxK

U2 - 10.1107/S1744309108038724

DO - 10.1107/S1744309108038724

M3 - Article

VL - 64

SP - 1184

EP - 1187

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

IS - 12

ER -