NMR-based detection of hydrogen/deuterium exchange in liposome-embedded membrane proteins.
Membrane proteins play key roles in biology. Determination of their structure in a membrane environment, however, is highly challenging. To address this challenge, we developed an approach that couples hydrogen/deuterium exchange of membrane proteins to rapid unfolding and detection by solution-state NMR spectroscopy. We show that the method allows analysis of the solvent protection of single residues in liposome-embedded proteins such as the 349-residue Tom40, the major protein translocation pore in the outer mitochondrial membrane, which has resisted structural analysis for many years.
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Xuejun
X
Yao
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Ulrich H N
UH
Dürr
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Zrinka
Z
Gattin
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Yvonne
Y
Laukat
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Rhagavendran L
RL
Narayanan
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Ann-Kathrin
AK
Brückner
Institut für Biochemie und Molekularbiologie, ZBMZ and BIOSS Centre for Biological Signalling Studies, Universität Freiburg, Freiburg, Germany.
Chris
C
Meisinger
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Adam
A
Lange
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Stefan
S
Becker
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany; German Center for Neurodegenerative Diseases (DZNE), Goöttingen, Germany; Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), University Medical Center, Göttingen, Germany.
Markus
M
Zweckstetter
D041961:Deuterium Exchange Measurement; D005656:Fungal Proteins / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D024101:Mitochondrial Proteins / Q000737:chemistry*; D009492:Neurospora crassa / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D058767:Protein Unfolding
D005656:Fungal Proteins; D008081:Liposomes; D008565:Membrane Proteins; D024101:Mitochondrial Proteins