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Purification, crystallization, and X-ray diffraction analysis of myocyte enhancer factor 2D and DNA complex by Chen, Zhiming; Wang, Qianqian; Zhang, Hao; Ma, Xiaodan; Wu, Wenyu; Cheng, Nuo; Zhang, Ji; Zhou, Aiwu; Li, Yuwen; Meng, Guoyu is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Purification, crystallization, and X-ray diffraction analysis of myocyte enhancer factor 2D and DNA complex about?

MEF2D-fusions have recently been identified as one of the major oncogenic drivers in precursor B-cell acute lymphoblastic leukemia (B-ALL). More importantly, they are often associated with patients with poor prognosis in B-ALL. To have a better understanding of the pathogenic mechanism underpinning MEF2D-fusions-driven leukemogenesis, it's essential to uncover the related structure information. In this study, we expressed and purified the MEF2D N-terminal DNA binding domain. The recombinant protein was engineered by cloning the encoding gene into the expression vector pET-32 m. A series of chromatographic steps involving affinity, ion-exchange and gel-filtration chromatography were used to achieve a final purity of >95%. For the crystallization of the MEF2D-DNA complex, a double-stranded DNA encoding 5'-AACTATTTATAAGA-3' and 5'-TTCTTATAAATAGT-3' was used (Wu et al., 2010) [1]. The MEF2D-DNA crystal with the size of about 20 μm × 20 μm × 20 μm was obtained at a final concentration of 12 mg/ml at the reservoir condition containing 30% PEG1500. The X-ray examination showed that the MEF2D-DNA crystal diffracted to 4.5 Å resolution, and belonged to space group P1, with unit-cell paramet

Who reads Purification, crystallization, and X-ray diffraction analysis of myocyte enhancer factor 2D and DNA complex?

Author

Chen, Zhiming; Wang, Qianqian; Zhang, Hao; Ma, Xiaodan; Wu, Wenyu; Cheng, Nuo; Zhang, Ji; Zhou, Aiwu; Li, Yuwen; Meng, Guoyu

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 1046-5928)

Published

2020

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)