In vitro folding and characterization of the p53 DNA binding domain

被引：5
作者：
Klein, C
Hesse, F
Dehner, A
Engh, RA
Schwaiger, M
Hansen, S
机构：
[1] Roche Diagnost GmbH, Pharma Res, D-82372 Penzberg, Germany
[2] Max Planck Inst Biochem, Abt Strukturforsch, D-82152 Martinsried, Germany
[3] Tech Univ Munich, Inst Organ Chem & Biochem, D-85747 Garching, Germany
来源：
BIOLOGICAL CHEMISTRY | 2004年 / 385卷 / 01期
关键词：
DNA binding; factorial screen; in vitro folding; NMR spectroscopy; p53;
D O I：
10.1515/BC.2004.012
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The transcription factor p53 acts as major tumor suppressor and is inactivated by mutation in more than 50% of all human tumors. We have established an efficient procedure for the in vitro folding and purification of the p53 DNA binding domain (p53DBD) using a modified factorial matrix approach that supplies large amounts of homogeneous (isotope-labeled) p53DBD for application in biochemical, crystallographic and NMR spectroscopic studies., We further show with biophysical methods that in vitro folded p53DBD is fully functional and that its conformation is identical to that obtained from the soluble fraction.
引用
收藏
页码：95 / 102
页数：8
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