High-resolution structure of a Na+/H+ antiporter dimer obtained by pulsed electron paramagnetic resonance distance measurements.
Transient or partial formation of complexes between biomacromolecules is a general mechanism used to control cellular functions. Several of these complexes escape structure determination by crystallographic means. We developed a new approach for determining the structure of protein dimers in the native environment (e.g., in the membrane) with high resolution in cases where the structure of the two monomers is known. The approach is based on measurements of distance distributions between spin labels in the range between 2 and 6 nanometers by a pulsed electron paramagnetic resonance technique and explicit modeling of spin label conformations. By applying this method to the membrane protein homodimer of the Na(+)/H(+) antiporter NhaA of Escherichia coli, the structure of the presumably physiological dimer was determined. It reveals two points of contact between the two monomers, with one of them confirming results of earlier cross-linking experiments.
Hilger
D
D
Ludwig-Maximilians-Universität München, Department Biologie I, D-80638 Munich, Germany.
Polyhach
Y
Y
Padan
E
E
Jung
H
H
Jeschke
G
G
D000483:Alleles; D001665:Binding Sites; D020285:Cryoelectron Microscopy; D003545:Cysteine / Q000737:chemistry; D030562:Databases, Protein; D019281:Dimerization; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015233:Models, Statistical; D008968:Molecular Conformation; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry* / Q000378:metabolism
D029968:Escherichia coli Proteins; C102839:NhaA protein, E coli; D017923:Sodium-Hydrogen Exchangers; D003545:Cysteine
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