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 R.
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
U2 - 10.1107/S1744309108038724
DO - 10.1107/S1744309108038724
M3 - Article
C2 - 19052383
AN - SCOPUS:57349105289
SN - 1744-3091
VL - 64
SP - 1184
EP - 1187
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 12
ER -