scholarly journals Simultaneous assignment and structure determination of a membrane protein from NMR orientational restraints

2003 ◽  
Vol 12 (3) ◽  
pp. 403-411 ◽  
Author(s):  
F. M. Marassi
Keyword(s):  
Membrane Protein ◽  
Structure Determination ◽  
Orientational Restraints

scholarly journals Structure determination of an integral membrane protein at room temperature from crystalsin situ

2015 ◽  
Vol 71 (6) ◽  
pp. 1228-1237 ◽  
Author(s):  
Danny Axford ◽  
James Foadi ◽  
Nien-Jen Hu ◽  
Hassanul Ghani Choudhury ◽  
So Iwata ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Structure Determination ◽  
Room Temperature ◽  
Protein Structures ◽  
Integral Membrane Protein ◽  
Data Sets ◽  
X Ray Diffraction ◽  
Haemophilus Influenza

The structure determination of an integral membrane protein using synchrotron X-ray diffraction data collected at room temperature directly in vapour-diffusion crystallization plates (in situ) is demonstrated. Exposing the crystalsin situeliminates manual sample handling and, since it is performed at room temperature, removes the complication of cryoprotection and potential structural anomalies induced by sample cryocooling. Essential to the method is the ability to limit radiation damage by recording a small amount of data per sample from many samples and subsequently assembling the resulting data sets using specialized software. The validity of this procedure is established by the structure determination ofHaemophilus influenzaTehA at 2.3 Å resolution. The method presented offers an effective protocol for the fast and efficient determination of membrane-protein structures at room temperature using third-generation synchrotron beamlines.

scholarly journals Membrane protein structure determination by SAD, SIR, or SIRAS phasing in serial femtosecond crystallography using an iododetergent

2016 ◽  
Vol 113 (46) ◽  
pp. 13039-13044 ◽  
Author(s):  
Takanori Nakane ◽  
Shinya Hanashima ◽  
Mamoru Suzuki ◽  
Haruka Saiki ◽  
Taichi Hayashi ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Structure Determination ◽  
Free Electron ◽  
Heavy Atom ◽  
Anomalous Scattering ◽  
X Ray ◽  
Isomorphous Replacement ◽  
Experimental Phasing ◽  
Serial Femtosecond Crystallography

The 3D structure determination of biological macromolecules by X-ray crystallography suffers from a phase problem: to perform Fourier transformation to calculate real space density maps, both intensities and phases of structure factors are necessary; however, measured diffraction patterns give only intensities. Although serial femtosecond crystallography (SFX) using X-ray free electron lasers (XFELs) has been steadily developed since 2009, experimental phasing still remains challenging. Here, using 7.0-keV (1.771 Å) X-ray pulses from the SPring-8 Angstrom Compact Free Electron Laser (SACLA), iodine single-wavelength anomalous diffraction (SAD), single isomorphous replacement (SIR), and single isomorphous replacement with anomalous scattering (SIRAS) phasing were performed in an SFX regime for a model membrane protein bacteriorhodopsin (bR). The crystals grown in bicelles were derivatized with an iodine-labeled detergent heavy-atom additive 13a (HAD13a), which contains the magic triangle, I3C head group with three iodine atoms. The alkyl tail was essential for binding of the detergent to the surface of bR. Strong anomalous and isomorphous difference signals from HAD13a enabled successful phasing using reflections up to 2.1-Å resolution from only 3,000 and 4,000 indexed images from native and derivative crystals, respectively. When more images were merged, structure solution was possible with data truncated at 3.3-Å resolution, which is the lowest resolution among the reported cases of SFX phasing. Moreover, preliminary SFX experiment showed that HAD13a successfully derivatized the G protein-coupled A2a adenosine receptor crystallized in lipidic cubic phases. These results pave the way for de novo structure determination of membrane proteins, which often diffract poorly, even with the brightest XFEL beams.

scholarly journals Structure Determination of a Membrane Protein in Proteoliposomes

2012 ◽  
Vol 134 (4) ◽  
pp. 2047-2056 ◽  
Author(s):  
Bibhuti B. Das ◽  
Henry J. Nothnagel ◽  
George J. Lu ◽  
Woo Sung Son ◽  
Ye Tian ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Structure Determination

Solid-state NMR spectroscopy structure determination of a lipid-embedded heptahelical membrane protein

2013 ◽  
Vol 10 (10) ◽  
pp. 1007-1012 ◽  
Author(s):  
Shenlin Wang ◽  
Rachel A Munro ◽  
Lichi Shi ◽  
Izuru Kawamura ◽  
Takashi Okitsu ◽  
...  
Keyword(s):  
Solid State ◽  
Nmr Spectroscopy ◽  
Membrane Protein ◽  
Structure Determination ◽  
Solid State Nmr ◽  
Solid State Nmr Spectroscopy

scholarly journals NMR Structure Determination of the Membrane Protein MerF in Bilayers

2012 ◽  
Vol 102 (3) ◽  
pp. 263a
Author(s):  
George J. Lu ◽  
Bibhuti B. Das ◽  
Henry J. Nothnagel ◽  
Woo Sung Son ◽  
Sang Ho Park ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Structure Determination ◽  
Nmr Structure ◽  
Nmr Structure Determination
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