Visualizing specific Cross-Protomer Interactions in the Homo-Oligomeric Membrane Protein Proteorhodopsin by DNP-enhanced Solid-state NMR.
J Am Chem Soc. 2015 Jun 23;
Authors: Maciejko J, Mehler M, Kaur J, Lieblein T, Morgner N, Ouari O, Tordo P, Becker-Baldus J, Glaubitz C
Abstract
Membrane proteins often form oligomeric complexes within the lipid bilayer but factors controlling their assembly are hard to predict and experimentally difficult to determine. An understanding of protein-protein interaction within the lipid bilayer is however required in order to elucidate the role of oligomerization for their functional mechanism and stabilization. Here, we demonstrate for the pentameric, heptahelical membrane protein green proteorhodopsin that solid-state NMR could identify specific interactions at the protomer interfaces, if the sensitivity is enhanced by dynamic nuclear polarization. For this purpose, differently labeled protomers have been assembled into the full pentamer complex embedded within the lipid bilayer. We show for this proof of concept that one specific salt bridge determines the formation of pentamers or hexamers. Data are supported by LILBID mass spectrometry and by Blue-native PAGE analysis. The presented approach is universally applicable and opens the door towards analyzing membrane protein interactions within homo-oligomers directly in the membrane.
PMID: 26102160 [PubMed - as supplied by publisher]
[NMR paper] Systematic solution to homo-oligomeric structures determined by NMR.
Systematic solution to homo-oligomeric structures determined by NMR.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--media.wiley.com-assets-2250-98-WileyOnlineLibrary-Button_120x27px_FullText.gif Related Articles Systematic solution to homo-oligomeric structures determined by NMR.
Proteins. 2015 Jan 23;
Authors: Martin JW, Zhou P, Donald BR
Abstract
Protein structure determination by NMR has predominantly relied on simulated annealing-based conformational search for a converged fold using primarily...
nmrlearner
Journal club
0
01-28-2015 03:22 AM
Direct Evidence of Imino Acid–Aromatic Interactions in Native Collagen Protein by DNP-Enhanced Solid-State NMR Spectroscopy
From The DNP-NMR Blog:
Direct Evidence of Imino Acid–Aromatic Interactions in Native Collagen Protein by DNP-Enhanced Solid-State NMR Spectroscopy
Singh, C., et al., Direct Evidence of Imino Acid–Aromatic Interactions in Native Collagen Protein by DNP-Enhanced Solid-State NMR Spectroscopy. The Journal of Physical Chemistry Letters, 2014. 5(22): p. 4044-4048.
http://dx.doi.org/10.1021/jz502081j
nmrlearner
News from NMR blogs
0
11-24-2014 04:15 PM
[NMR paper] High-resolution paramagnetically enhanced solid-state NMR spectroscopy of membrane proteins at fast magic angle spinning.
High-resolution paramagnetically enhanced solid-state NMR spectroscopy of membrane proteins at fast magic angle spinning.
Related Articles High-resolution paramagnetically enhanced solid-state NMR spectroscopy of membrane proteins at fast magic angle spinning.
J Biomol NMR. 2013 Dec 13;
Authors: Ward ME, Wang S, Krishnamurthy S, Hutchins H, Fey M, Brown LS, Ladizhansky V
Abstract
Magic angle spinning nuclear magnetic resonance (MAS NMR) is well suited for the study of membrane proteins in membrane mimetic and native membrane...
nmrlearner
Journal club
0
12-18-2013 04:00 PM
Solid-state magic-angle spinning NMR of membrane proteins and protein–ligand interactions
Solid-state magic-angle spinning NMR of membrane proteins and protein–ligand interactions
April 2012
Publication year: 2012
Source:European Journal of Cell Biology, Volume 91, Issue 4</br>
</br>
Structural biology is developing into a universal tool for visualizing biological processes in space and time at atomic resolution. The field has been built by established methodology like X-ray crystallography, electron microscopy and solution NMR and is now incorporating new techniques, such as small-angle X-ray scattering, electron tomography, magic-angle-spinning solid-state...
nmrlearner
Journal club
0
02-03-2013 10:13 AM
Proton-Detected Solid-State NMR Reveals Intramembrane Polar Networks in a Seven-Helical Transmembrane Protein Proteorhodopsin
Proton-Detected Solid-State NMR Reveals Intramembrane Polar Networks in a Seven-Helical Transmembrane Protein Proteorhodopsin
Meaghan E. Ward, Lichi Shi, Evelyn Lake, Sridevi Krishnamurthy, Howard Hutchins, Leonid S. Brown and Vladimir Ladizhansky
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja207137h/aop/images/medium/ja-2011-07137h_0008.gif
Journal of the American Chemical Society
DOI: 10.1021/ja207137h
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/Vzwkh1cjxOU
nmrlearner
Journal club
0
10-09-2011 06:15 AM
Proton Detected Solid-State NMR Reveals Intramembrane Polar Networks in a Seven-Helical Transmembrane Protein Proteorhodopsin.
Proton Detected Solid-State NMR Reveals Intramembrane Polar Networks in a Seven-Helical Transmembrane Protein Proteorhodopsin.
Proton Detected Solid-State NMR Reveals Intramembrane Polar Networks in a Seven-Helical Transmembrane Protein Proteorhodopsin.
J Am Chem Soc. 2011 Sep 16;
Authors: Ward ME, Shi L, Lake EM, Krishnamurthy S, Hutchins H, Brown LS, Ladizhansky V
Abstract
We used high-resolution proton-detected multidimensional NMR to study the solvent-exposed parts of an integral seven-helical membrane proton pump proteorhodopsin...
nmrlearner
Journal club
0
09-17-2011 08:21 PM
Solid-state NMR of amyloid membrane interactions.
Solid-state NMR of amyloid membrane interactions.
Solid-state NMR of amyloid membrane interactions.
Methods Mol Biol. 2011;752:165-77
Authors: Gehman JD, Separovic F
Solid-state NMR pulse sequences often feature fewer pulses and delays than the more common solution NMR experiments. This ostensible simplicity, however, belies the care with which experimental parameters must be determined, as solid-state NMR can be much less forgiving of improper experimental set-up. This is especially true of "semi-solid" samples, such as the phospholipid vesicles...
nmrlearner
Journal club
0
06-30-2011 01:24 PM
[NMR paper] Synthesis, enhanced fusogenicity, and solid state NMR measurements of cross-linked HI
Synthesis, enhanced fusogenicity, and solid state NMR measurements of cross-linked HIV-1 fusion peptides.
Related Articles Synthesis, enhanced fusogenicity, and solid state NMR measurements of cross-linked HIV-1 fusion peptides.
Biochemistry. 2003 Apr 1;42(12):3527-35
Authors: Yang R, Yang J, Weliky DP
In the HIV-1 gp41 and other viral fusion proteins, the minimal oligomerization state is believed to be trimeric with three N-terminal fusion peptides inserting into the membrane in close proximity. Previous studies have demonstrated that the...
Visualizing specific Cross-Protomer Interactions in the Homo-Oligomeric Membrane Protein Proteorhodopsin by DNP-enhanced Solid-state NMR.
Linear Mode
