Detection of membrane protein two-dimensional crystals in living cells

E. J. Gualtieri, F. Guo, D. J. Kissick, J. Jose, R. J. Kuhn, W. Jiang, G. J. Simpson

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

It is notoriously difficult to grow membrane protein crystals and solve membrane protein structures. Improved detection and screening of membrane protein crystals are needed. We have shown here that second-order nonlinear optical imaging of chiral crystals based on second harmonic generation can provide sensitive and selective detection of twodimensional protein crystalline arrays with sufficiently low background to enable crystal detection within the membranes of live cells. The method was validated using bacteriorhodopsin crystals generated in live Halobacterium halobium bacteria and confirmed by electron microscopy from the isolated crystals. Additional studies of alphavirus glycoproteins indicated the presence of localized crystalline domains associated with virus budding from mammalian cells. These results suggest that in vivo crystallization may provide a means for expediting membrane protein structure determination for proteins exhibiting propensities for two-dimensional crystal formation.

Original languageEnglish (US)
Pages (from-to)207-214
Number of pages8
JournalBiophysical journal
Volume100
Issue number1
DOIs
StatePublished - Jan 5 2011

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Membrane Proteins
Halobacterium salinarum
Alphavirus
Virus Release
Satellite Viruses
Bacteriorhodopsins
Protein Array Analysis
Optical Imaging
Crystallization
Electron Microscopy
Glycoproteins
Cell Membrane
Bacteria
Proteins

All Science Journal Classification (ASJC) codes

  • Biophysics

Cite this

Gualtieri, E. J., Guo, F., Kissick, D. J., Jose, J., Kuhn, R. J., Jiang, W., & Simpson, G. J. (2011). Detection of membrane protein two-dimensional crystals in living cells. Biophysical journal, 100(1), 207-214. https://doi.org/10.1016/j.bpj.2010.10.051
Gualtieri, E. J. ; Guo, F. ; Kissick, D. J. ; Jose, J. ; Kuhn, R. J. ; Jiang, W. ; Simpson, G. J. / Detection of membrane protein two-dimensional crystals in living cells. In: Biophysical journal. 2011 ; Vol. 100, No. 1. pp. 207-214.
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Gualtieri, EJ, Guo, F, Kissick, DJ, Jose, J, Kuhn, RJ, Jiang, W & Simpson, GJ 2011, 'Detection of membrane protein two-dimensional crystals in living cells', Biophysical journal, vol. 100, no. 1, pp. 207-214. https://doi.org/10.1016/j.bpj.2010.10.051

Detection of membrane protein two-dimensional crystals in living cells. / Gualtieri, E. J.; Guo, F.; Kissick, D. J.; Jose, J.; Kuhn, R. J.; Jiang, W.; Simpson, G. J.

In: Biophysical journal, Vol. 100, No. 1, 05.01.2011, p. 207-214.

Research output: Contribution to journalArticle

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