| Autor: |
Huang, X, Crute, B E, Sun, C, Tang, Y Y, Kelley, J J, Lewis, A F, Hartman, K L, Laue, T M, Speck, N A, Bushweller, J H |
| Zdroj: |
Journal of Biological Chemistry; January 1998, Vol. 273 Issue: 4 p2480-2487, 8p |
| Abstrakt: |
Core-binding factors (CBF) are heteromeric transcription factors essential for several developmental processes, including hematopoiesis. CBFs contain a DNA-binding CBF alpha subunit and a non-DNA binding CBF beta subunit that increases the affinity of CBF alpha for DNA. We have developed a procedure for overexpressing and purifying full-length CBF beta as well as a truncated form containing the N-terminal 141 amino acids using a novel glutaredoxin fusion expression system. Substantial quantities of the CBF beta proteins can be produced in this manner allowing for their biophysical characterization. We show that the full-length and truncated forms of CBF beta bind to a CBF alpha DNA complex with very similar affinities. Sedimentation equilibrium measurements show these proteins to be monomeric. Circular dichroism spectroscopy demonstrates that CBF beta is a mixed alpha/beta protein and NMR spectroscopy shows that the truncated and full-length proteins are structurally similar and suitable for structure determination by NMR spectroscopy. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
