7 Å projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals

Julie E. Norville, Deb Kelly, Thomas F. Knight, Angela M. Belcher, Thomas Walz

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Two-dimensional crystallization on lipid monolayers is a versatile tool to obtain structural information of proteins by electron microscopy. An inherent problem with this approach is to prepare samples in a way that preserves the crystalline order of the protein array and produces specimens that are sufficiently flat for high-resolution data collection at high tilt angles. As a test specimen to optimize the preparation of lipid monolayer crystals for electron microscopy imaging, we used the S-layer protein sbpA, a protein with potential for designing arrays of both biological and inorganic materials with engineered properties for a variety of nanotechnology applications. Sugar embedding is currently considered the best method to prepare two-dimensional crystals of membrane proteins reconstituted into lipid bilayers. We found that using a loop to transfer lipid monolayer crystals to an electron microscopy grid followed by embedding in trehalose and quick-freezing in liquid ethane also yielded the highest resolution images for sbpA lipid monolayer crystals. Using images of specimens prepared in this way we could calculate a projection map of sbpA at 7 Å resolution, one of the highest resolution projection structures obtained with lipid monolayer crystals to date.

Original languageEnglish (US)
Pages (from-to)313-323
Number of pages11
JournalJournal of Structural Biology
Volume160
Issue number3
DOIs
StatePublished - Dec 1 2007

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Trehalose
Lipids
Electron Microscopy
Protein Array Analysis
Nanotechnology
Ethane
Lipid Bilayers
Crystallization
Freezing
Membrane Proteins
Proteins
S-layer proteins

All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Norville, Julie E. ; Kelly, Deb ; Knight, Thomas F. ; Belcher, Angela M. ; Walz, Thomas. / 7 Å projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals. In: Journal of Structural Biology. 2007 ; Vol. 160, No. 3. pp. 313-323.
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7 Å projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals. / Norville, Julie E.; Kelly, Deb; Knight, Thomas F.; Belcher, Angela M.; Walz, Thomas.

In: Journal of Structural Biology, Vol. 160, No. 3, 01.12.2007, p. 313-323.

Research output: Contribution to journalArticle

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