Abstract

Research Article1 October 1990free access Protein–DNA contacts in the structure of a homeodomain–DNA complex determined by nuclear magnetic resonance spectroscopy in solution. G. Otting G. Otting Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author Y. Q. Qian Y. Q. Qian Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Billeter M. Billeter Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Müller M. Müller Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Affolter M. Affolter Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author W. J. Gehring W. J. Gehring Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author K. Wüthrich K. Wüthrich Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author G. Otting G. Otting Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author Y. Q. Qian Y. Q. Qian Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Billeter M. Billeter Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Müller M. Müller Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author M. Affolter M. Affolter Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author W. J. Gehring W. J. Gehring Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author K. Wüthrich K. Wüthrich Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. Search for more papers by this author Author Information G. Otting1, Y. Q. Qian1, M. Billeter1, M. Müller1, M. Affolter1, W. J. Gehring1 and K. Wüthrich1 1Institut für Molekularbiologie und Biophysik. ETH-Hönggerberg, Zürich, Switzerland. The EMBO Journal (1990)9:3085-3092https://doi.org/10.1002/j.1460-2075.1990.tb07505.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info The 1:1 complex of the mutant Antp(C39––S) homeodomain with a 14 bp DNA fragment corresponding to the BS2 binding site was studied by nuclear magnetic resonance (NMR) spectroscopy in aqueous solution. The complex has a molecular weight of 17,800 and its lifetime is long compared with the NMR chemical shift time scale. Investigations of the three-dimensional structure were based on the use of the fully 15N-labelled protein, two-dimensional homonuclear proton NOESY with 15N(omega 2) half-filter, and heteronuclear three-dimensional NMR experiments. Based on nearly complete sequence-specific resonance assignments, both the protein and the DNA were found to have similar conformations in the free form and in the complex. A sufficient number of intermolecular 1H-1H Overhauser effects (NOE) could be identified to enable a unique docking of the protein on the DNA, which was achieved with the use of an ellipsoid algorithm. In the complex there are intermolecular NOEs between the elongated second helix in the helix-turn-helix motif of the homeodomain and the major groove of the DNA. Additional NOE contacts with the DNA involve the polypeptide loop immediately preceding the helix-turn-helix segment, and Arg5. This latter contact is of special interest, both because Arg5 reaches into the minor groove and because in the free Antp(C39––S) homeodomain no defined spatial structure could be found for the apparently flexible N-terminal segment comprising residues 0-6. Previous ArticleNext Article Volume 9Issue 101 October 1990In this issue RelatedDetailsLoading ...