X-ray diffraction from Membrane protein nanocrystals

M. S. Hunter, D. P. DePonte, D. A. Shapiro, Richard Kirian, X. Wang, D. Starodub, S. Marchesini, Uwe Weierstall, R. B. Doak, John Spence, Petra Fromme

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Membrane proteins constitute >30% of the proteins in an average cell, and yet the number of currently known structures of unique membrane proteins is <300. To develop new concepts for membrane protein structure determination, we have explored the serial nanocrystallography method, in which fully hydrated protein nanocrystals are delivered to an x-ray beam within a liquid jet at room temperature. As a model system, we have collected x-ray powder diffraction data from the integral membrane protein Photosystem I, which consists of 36 subunits and 381 cofactors. Data were collected from crystals ranging in size from 100 nm to 2 &mu;m. The results demonstrate that there are membrane protein crystals that contain <100 unit cells (200 total molecules) and that 3D crystals of membrane proteins, which contain <200 molecules, may be suitable for structural investigation. Serial nanocrystallography overcomes the problem of x-ray damage, which is currently one of the major limitations for x-ray structure determination of small crystals. By combining serial nanocrystallography with x-ray free-electron laser sources in the future, it may be possible to produce molecular-resolution electron-density maps using membrane protein crystals that contain only a few hundred or thousand unit cells.

Original languageEnglish (US)
Pages (from-to)198-206
Number of pages9
JournalBiophysical Journal
Volume100
Issue number1
DOIs
StatePublished - Jan 5 2011

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X-Ray Diffraction
Nanoparticles
Membrane Proteins
X-Rays
Electrons
Photosystem I Protein Complex
Powder Diffraction
Proteins
Lasers
Cell Count
Temperature

ASJC Scopus subject areas

  • Biophysics

Cite this

Hunter, M. S., DePonte, D. P., Shapiro, D. A., Kirian, R., Wang, X., Starodub, D., ... Fromme, P. (2011). X-ray diffraction from Membrane protein nanocrystals. Biophysical Journal, 100(1), 198-206. https://doi.org/10.1016/j.bpj.2010.10.049

X-ray diffraction from Membrane protein nanocrystals. / Hunter, M. S.; DePonte, D. P.; Shapiro, D. A.; Kirian, Richard; Wang, X.; Starodub, D.; Marchesini, S.; Weierstall, Uwe; Doak, R. B.; Spence, John; Fromme, Petra.

In: Biophysical Journal, Vol. 100, No. 1, 05.01.2011, p. 198-206.

Research output: Contribution to journalArticle

Hunter, MS, DePonte, DP, Shapiro, DA, Kirian, R, Wang, X, Starodub, D, Marchesini, S, Weierstall, U, Doak, RB, Spence, J & Fromme, P 2011, 'X-ray diffraction from Membrane protein nanocrystals', Biophysical Journal, vol. 100, no. 1, pp. 198-206. https://doi.org/10.1016/j.bpj.2010.10.049
Hunter MS, DePonte DP, Shapiro DA, Kirian R, Wang X, Starodub D et al. X-ray diffraction from Membrane protein nanocrystals. Biophysical Journal. 2011 Jan 5;100(1):198-206. https://doi.org/10.1016/j.bpj.2010.10.049
Hunter, M. S. ; DePonte, D. P. ; Shapiro, D. A. ; Kirian, Richard ; Wang, X. ; Starodub, D. ; Marchesini, S. ; Weierstall, Uwe ; Doak, R. B. ; Spence, John ; Fromme, Petra. / X-ray diffraction from Membrane protein nanocrystals. In: Biophysical Journal. 2011 ; Vol. 100, No. 1. pp. 198-206.
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