Mapping structural interactions using in-cell NMR spectroscopy (STINT-NMR)

Burz, David S.; Dutta, Kaushik; Cowburn, David; Shekhtman, Alexander
February 2006
Nature Methods;Feb2006, Vol. 3 Issue 2, p91
Academic Journal
We describe a high-throughput in-cell nuclear magnetic resonance (NMR)-based method for mapping the structural changes that accompany protein-protein interactions (STINT-NMR). The method entails sequentially expressing two (or more) proteins within a single bacterial cell in a time-controlled manner and monitoring the protein interactions using in-cell NMR spectroscopy. The resulting spectra provide a complete titration of the interaction and define structural details of the interacting surfaces at atomic resolution.


Related Articles

  • Optimized set of two-dimensional experiments for fast sequential assignment, secondary structure determination, and backbone fold validation of 13C/15N-labelled proteins. Bersch, Beate; Rossy, Emmanuel; Cov�s, Jacques; Brutscher, Bernhard // Journal of Biomolecular NMR;Sep2003, Vol. 27 Issue 1, p57 

    NMR experiments are presented which allow backbone resonance assignment, secondary structure identification, and in favorable cases also molecular fold topology determination from a series of two-dimensional 1H-15N HSQC-like spectra. The 1H-15N correlation peaks are frequency shifted by an...

  • Fully automated sequence-specific resonance assignments of hetero- nuclear protein spectra. Malmodin, Daniel; Papavoine, Christina H. M.; Billeter, Martin // Journal of Biomolecular NMR;Sep2003, Vol. 27 Issue 1, p69 

    Full automation of the analysis of spectra is a prerequisite for high-throughput NMR studies in structural or functional genomics. Sequence-specific assignments often form the major bottleneck. Here, we present a procedure that yields nearly complete backbone and side chain resonance assignments...

  • Validation of the GROMOS force-field parameter set 45A3 against nuclear magnetic resonance data of hen egg lysozyme. Soares, T. A.; Daura, X.; Oostenbrink, C.; Smith, L. J.; van Gunsteren, W. F. // Journal of Biomolecular NMR;Dec2004, Vol. 30 Issue 4, p407 

    The quality of molecular dynamics (MD) simulations of proteins depends critically on the biomolecular force field that is used. Such force fields are defined by force-field parameter sets, which are generally determined and improved through calibration of properties of small molecules against...

  • Structural biology: Designer labels. Opella, Stanley J. // Nature;3/2/2006, Vol. 440 Issue 7080, p40 

    The article discusses on several research studies related to the use of the nuclear magnetic resonance (NMR) spectroscopy in determining of the structures of larger proteins. It can be used on molecules in all physical states and degrees or order, and is capable of determining the structures of...

  • NMR studies of structure and function of biological macromolecules (Nobel Lecture)*. W�thrich, Kurt // Journal of Biomolecular NMR;Sep2003, Vol. 27 Issue 1, p13 

    The article presents nuclear magnetic resonance (NMR) studies of structure and function of biological macromolecules. NMR spectroscopy is unique among the methods available for three-dimensional structure determination of proteins and nucleic acids at atomic resolution, since the NMR data can be...

  • GETTING TO GRIPS WITH HA-PROTEIN INTERACTIONS. Blundell, Charles D.; Kahmann, Jan D.; Perczel, András; Mahoney, David J.; Cordell, Martin R.; Teriete, Peter; Campbell, Iain D.; Day, Anthony J. // Hyaluronan;2002, Vol. 1, p161 

    The interactions between HA and proteins are most commonly mediated by a domain termed a Link module. The Link module from human TSG-6, produced by expression in E. coli, has been used previously to determine its tertiary structure and identify the position of the HA-binding site by NMR...

  • Direct detection of CH/Ï€ interactions in proteins. Plevin, Michael J.; Bryce, David L.; Boisbouvier, Jérôme // Nature Chemistry;Jun2010, Vol. 2 Issue 6, p466 

    XH/π interactions make important contributions to biomolecular structure and function. These weakly polar interactions, involving π-system acceptor groups, are usually identified from the three-dimensional structures of proteins. Here, nuclear magnetic resonance spectroscopy has been...

  • Magnetic resonance in the solid state: applications to protein folding, amyloid fibrils and membrane proteins. Baldus, Marc // European Biophysics Journal;Jun2007 Supplement, Vol. 36, p37 

    Solid-state nuclear magnetic resonance (ssNMR) represents a spectroscopic method to study non-crystalline molecules at atomic resolution. Advancements in spectroscopy and biochemistry provide increasing possibilities to study structure and dynamics of complex biomolecular systems by ssNMR. Here,...

  • RefDB: A database of uniformly referenced protein chemical shifts. Zhang, Haiyan; Neal, Stephen; Wishart, David S. // Journal of Biomolecular NMR;Mar2003, Vol. 25 Issue 3, p173 

    RefDB is a secondary database of reference-corrected protein chemical shifts derived from the BioMagResBank (BMRB). The database was assembled by using a recently developed program (SHIFTX) to predict protein 1H, 13C and 15N chemical shifts from X-ray or NMR coordinate data of previously...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics