Department
Chemistry and Biochemistry
Faculty Advisor
Dr. Fenfei Lang
Location
East and Center Ballrooms
Start Date
17-3-2015 3:00 PM
End Date
17-3-2015 4:00 PM
Session
Session 3
Session Topic
Poster
Abstract
The mammalian high mobility group protein AT-hook 2 (HMGA2) is a nuclear oncoprotein, and its abnormal expression is directly linked to tumorigenesis. The HMGA2 gene is located on the chromosomal bands 12q13-15. Rearrangement of these bands disrupts the normal functions of HMGA2 and causes the formation of benign tumors, such as lipomas, and malignant tumors, such as lung cancer. In this study, our objective was to make sufficient amounts of 15N-, 13C-labeled HMGA2 for NMR studies to obtain a high-resolution structure of HMGA2-DNA complexes. In order to label this protein, E. coli strain BLR(DE3), containing a plasmid expressing HMGA2, was grown in minimal M9 medium containing 15NH4Cl and 13C-glucose. The 15N-, 13Clabeled HMGA2 protein was purified according to a purification protocol previously published by our research group. SDS-PAGE, Western Blotting, and Mass Spectrometry were used to demonstrate the successful expression and purification of the 15N-, 13C-labeled HMGA2. HMGA2 protein samples were sent to our collaborators at the University of Bayreuth, Germany for NMR structural studies.
File Type
Poster
Expression and Purification of 15N, 13Clabeled HMGA2 for NMR Structural Studies
East and Center Ballrooms
The mammalian high mobility group protein AT-hook 2 (HMGA2) is a nuclear oncoprotein, and its abnormal expression is directly linked to tumorigenesis. The HMGA2 gene is located on the chromosomal bands 12q13-15. Rearrangement of these bands disrupts the normal functions of HMGA2 and causes the formation of benign tumors, such as lipomas, and malignant tumors, such as lung cancer. In this study, our objective was to make sufficient amounts of 15N-, 13C-labeled HMGA2 for NMR studies to obtain a high-resolution structure of HMGA2-DNA complexes. In order to label this protein, E. coli strain BLR(DE3), containing a plasmid expressing HMGA2, was grown in minimal M9 medium containing 15NH4Cl and 13C-glucose. The 15N-, 13Clabeled HMGA2 protein was purified according to a purification protocol previously published by our research group. SDS-PAGE, Western Blotting, and Mass Spectrometry were used to demonstrate the successful expression and purification of the 15N-, 13C-labeled HMGA2. HMGA2 protein samples were sent to our collaborators at the University of Bayreuth, Germany for NMR structural studies.
Rights Statement
In Copyright. URI: http://rightsstatements.org/vocab/InC/1.0/
This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
Comments
**Abstract Only**