Abstract
We describe a procedure to obtain structural data of biological macromolecules from small 2-D crystals using cryo-electron crystallography. The procedure has been applied to a membrane-embedded multi-protein complex (photosystem II). This complex, under certain conditions, forms 2-D crystals in its native membrane which are composed of many small mosaic patches and therefore represents an ideal test specimen. By averaging in reciprocal space over more than 50 crystalline patches, it was possible to calculate a projection map using data approaching 1.3 nm resolution while in the Fourier transforms of individual patches only reflections up to the 4th order were readily identifiable. The efficiency of the procedure was evaluated by examining the fidelity of (i) the rotational alignment of patches prior to merging, (ii) the space group assignment, (iii) phase and amplitude extraction, and (iv) the correction of the structure factors for the effects of the contrast transfer function (CTF).
| Original language | English (US) |
|---|---|
| Pages (from-to) | 113-128 |
| Number of pages | 16 |
| Journal | Ultramicroscopy |
| Volume | 77 |
| Issue number | 3-4 |
| DOIs | |
| State | Published - Jul 1999 |
| Externally published | Yes |
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Keywords
- Cryo-electron crystallography
- Image processing
- Mosaicity
- Small 2-D crystals
ASJC Scopus subject areas
- Materials Science(all)
- Instrumentation
Cite this
Cryo-electron crystallography of small and mosaic 2-D crystals : An assessment of a procedure for high-resolution data retrieval. / Stoylova, Svetla S.; Ford, Robert C.; Holzenburg, Andreas.
In: Ultramicroscopy, Vol. 77, No. 3-4, 07.1999, p. 113-128.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cryo-electron crystallography of small and mosaic 2-D crystals
T2 - An assessment of a procedure for high-resolution data retrieval
AU - Stoylova, Svetla S.
AU - Ford, Robert C.
AU - Holzenburg, Andreas
PY - 1999/7
Y1 - 1999/7
N2 - We describe a procedure to obtain structural data of biological macromolecules from small 2-D crystals using cryo-electron crystallography. The procedure has been applied to a membrane-embedded multi-protein complex (photosystem II). This complex, under certain conditions, forms 2-D crystals in its native membrane which are composed of many small mosaic patches and therefore represents an ideal test specimen. By averaging in reciprocal space over more than 50 crystalline patches, it was possible to calculate a projection map using data approaching 1.3 nm resolution while in the Fourier transforms of individual patches only reflections up to the 4th order were readily identifiable. The efficiency of the procedure was evaluated by examining the fidelity of (i) the rotational alignment of patches prior to merging, (ii) the space group assignment, (iii) phase and amplitude extraction, and (iv) the correction of the structure factors for the effects of the contrast transfer function (CTF).
AB - We describe a procedure to obtain structural data of biological macromolecules from small 2-D crystals using cryo-electron crystallography. The procedure has been applied to a membrane-embedded multi-protein complex (photosystem II). This complex, under certain conditions, forms 2-D crystals in its native membrane which are composed of many small mosaic patches and therefore represents an ideal test specimen. By averaging in reciprocal space over more than 50 crystalline patches, it was possible to calculate a projection map using data approaching 1.3 nm resolution while in the Fourier transforms of individual patches only reflections up to the 4th order were readily identifiable. The efficiency of the procedure was evaluated by examining the fidelity of (i) the rotational alignment of patches prior to merging, (ii) the space group assignment, (iii) phase and amplitude extraction, and (iv) the correction of the structure factors for the effects of the contrast transfer function (CTF).
KW - Cryo-electron crystallography
KW - Image processing
KW - Mosaicity
KW - Small 2-D crystals
UR - http://www.scopus.com/inward/record.url?scp=0033166554&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033166554&partnerID=8YFLogxK
U2 - 10.1016/S0304-3991(99)00039-X
DO - 10.1016/S0304-3991(99)00039-X
M3 - Article
VL - 77
SP - 113
EP - 128
JO - Ultramicroscopy
JF - Ultramicroscopy
SN - 0304-3991
IS - 3-4
ER -