ABSTRACT A new principle in constructing molecular complexes from the known high-resolution domain structures joining data from NMR and small-angle x-ray scattering (SAXS) measurements is described. Structure of calmodulin in complex with trifluoperazine was built from N- and C-terminal domains oriented based on residual dipolar couplings measured by NMR in a dilute liquid crystal, and the overall shape of the complex was derived from SAXS data. The residual dipolar coupling data serves to reduce angular degrees of freedom, and the small-angle scattering data serves to confine the translational degrees of freedom. The complex built by this method was found to be consistent with the known crystal structure. The study demonstrates how approximate tertiary structures of modular proteins or quaternary structures composed of subunits can be assembled from high-resolution structures of domains or subunits using mutually complementary NMR and SAXS data.
We are indebted to Andrea Hounslow and Clare Treritt for advice and assistance.
This work was supported by the Academy of Finland and Technology Agent of Finland (TEKES).
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Maija-Liisa Mattinen,* Kimmo Paakkonen,* Teemu Ikonen,^ Jeremy Craven,^^ Torbjorn Drakenberg,* Ritva Serimaa,^ Jonathan Waltho,^^ and Arto Annila^
*VTT Biotechnology, FIN-02044 VTT, Espoo, Finland; ^Department of Physical Sciences, University of Helsinki, FIN-00014 Helsinki, Finland; and ^^Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom
Submitted October 31, 2001, and accepted for publication February 19, 2002.
Address reprint requests to Dr. Arto Annila, Institute of Biotechnology, University of Helsinki, P.O. Box 56, FIN-00014 Helsinki, Finland. Tel.: 358-9-191-50629; Fax: 358-9-191-50639; E-mail: firstname.lastname@example.org.
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