scholarly journals Selective Interface Detection:  Mapping Binding Site Contacts in Membrane Proteins by NMR Spectroscopy

2005 ◽  
Vol 127 (16) ◽  
pp. 5734-5735 ◽  
Author(s):  
Suzanne R. Kiihne ◽  
Alain F. L. Creemers ◽  
Willem J. de Grip ◽  
Petra H. M. Bovee-Geurts ◽  
Johan Lugtenburg ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Membrane Proteins ◽  
Binding Site

scholarly journals Monitoring Dynamics of Large Membrane Proteins by 19F Paramagnetic Longitudinal Relaxation: Domain Movement in a Glutamate Transporter Homolog

2019 ◽  
Author(s):  
Yun Huang ◽  
Xiaoyu Wang ◽  
Guohua Lv ◽  
Asghar M. Razavi ◽  
Gerard H. M. Huysmans ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Membrane Proteins ◽  
Binding Site ◽  
Conformational Changes ◽  
Glutamate Transporter ◽  
Conformational Exchange ◽  
Substrate Binding Site ◽  
Domain Movement ◽  
Paramagnetic Metal ◽  
Longitudinal Relaxation

AbstractIn proteins where conformational changes are functionally important, the number of accessible states and their dynamics are often difficult to establish. Here we describe a novel 19F-NMR spectroscopy approach to probe dynamics of large membrane proteins. We labeled a glutamate transporter homologue with a 19F probe via cysteine chemistry and with a Ni2+ ion via chelation by a di-histidine motif. We used distance-dependent enhancement of the longitudinal relaxation of 19F nuclei by the paramagnetic metal to assign the observed resonances. We identified two outward- and one inward-facing states of the transporter, in which the substrate-binding site is near the extracellular and intracellular solutions, respectively. We then resolved the structure of the unanticipated second outward-facing state by Cryo-EM. Finally, we showed that the rates of the conformational exchange are accessible from measurements of the metal-enhanced longitudinal relaxation of 19F nuclei.

Proton-Detected NMR Spectroscopy of Nanodisc-Embedded Membrane Proteins: MAS Solid-State vs Solution-State Methods

2017 ◽  
Vol 121 (32) ◽  
pp. 7671-7680 ◽  
Author(s):  
Nils-Alexander Lakomek ◽  
Lukas Frey ◽  
Stefan Bibow ◽  
Anja Böckmann ◽  
Roland Riek ◽  
...  
Keyword(s):  
Solid State ◽  
Nmr Spectroscopy ◽  
Membrane Proteins ◽  
Solution State

A Solid-State NMR Approach to Structure Determination of Membrane-Associated Peptides and Proteins

1997 ◽  
Author(s):  
S.J. Opella ◽  
L.E. Chirlian
Keyword(s):  
Nmr Spectroscopy ◽  
Membrane Proteins ◽  
Structural Biology ◽  
Experimental Studies ◽  
Globular Proteins ◽  
Biological Properties ◽  
Cellular Functions ◽  
X Ray ◽  
X Ray Crystallography ◽  
Form And Function

Structural biology relies on detailed descriptions of the three-dimensional structures of peptides, proteins, and other biopolymers to explain the form and function of biological systems ranging in complexity from individual molecules to entire organisms. NMR spectroscopy and X-ray crystallography, in combination with several types of calculations, provide the required structural information. In recent years, the structures of several hundred proteins have been determined by one or both of these experimental methods. However, since the protein molecules must either reorient rapidly in samples for multidimensional solution NMR spectroscopy or form high quality single crystals in samples for X-ray crystallography, nearly all of the structures determined up to now have been of the soluble, globular proteins that are found in the cytoplasm and periplasmof cells and fortuitously have these favorable properties. Since only a minority of biological properties are expressed by globular proteins, and proteins, in general, have evolved in order to express specific functions rather than act as samples for experimental studies, there are other classes of proteins whose structures are currently unknown but are of keen interest in structural biology. More than half of all proteins appear to be associated with membranes, and many cellular functions are expressed by proteins in other types of supramolecular complexes with nucleic acids, carbohydrates, or other proteins. The interest in the structures of membrane proteins, structural proteins, and proteins in complexes provides many opportunities for the further development and application of NMR spectroscopy. Our understanding of polypeptides associated with lipids in membranes, in particular, is primitive, especially compared to that for globular proteins. This is largely a consequence of the experimental difficulties encountered in their study by conventional NMR and X-ray approaches. Fortunately, the principal features of two major classes of membrane proteins have been identified from studies of several tractable examples. Bacteriorhodopsin (Henderson et al., 1990), the subunits of the photosynthetic reaction center (Deisenhofer et al., 1985), and filamentous bacteriophage coat proteins (Shon et al., 1991; McDonnell et al., 1993) have all been shown to have long transmembrane hydrophobic helices, shorter amphipathic bridging helices in the plane of the bilayers, both structured and mobile loops connecting the helices, and mobile N- and C-terminal regions.

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