Multidimensional NMR spectroscopy of DNA-binding proteins: structure and function of a transcription factor

• Published in: Toxicology Letters Vol. 82; pp. 577 - 589
• Main Authors: Hsu, Victor L., Jia, Xin, Kearns, David R.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Shannon Elsevier Ireland Ltd 01.12.1995; Amsterdam Elsevier Science
• Subjects: Amino Acid Sequence; Biological and medical sciences; DNA-binding proteins; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - physiology; Fundamental and applied biological sciences. Psychology; Interactions. Associations; Intermolecular phenomena; Magnetic Resonance Spectroscopy; Molecular biophysics; Molecular Sequence Data; NMR spectroscopy; Protein Structure, Secondary; Structure determination; Transcription factor 1; Transcription Factors - chemistry; Transcription Factors - physiology; NMR spectroscopy; Structure determination; Transcription factor 1; DNA-binding proteins; Virus; Phage SP01; Molecular structure; DNA binding protein; Molecular interaction; NMR spectrometry; Transcription factor; Myoviridae; Nucleic acid; Phage
• ISSN: 0378-4274; 1879-3169
• DOI: 10.1016/0378-4274(95)03585-0

The solution structure of a type II DNA-binding protein (DBPII), transcription factor 1 (TF1), has been determined using NMR spectroscopy. A multidimensional, heteronuclear strategy was employed to overcome assignment ambiguities due to resonance overlap and broadened crosspeaks. This approach involved the use of selectively deuteriated, 13C- and 15N-labeled samples and ‘isotopic heterodimers’ to distinguish between intra- and intermonomeric NOEs. A comparison with the crystal structure and NMR analysis of the E. coli HU protein suggests that other homologous proteins in this family will possess similar tertiary structures. This NMR strategy is applicable to the study of other proteins and their biomolecular complexes.