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3D Reconstruction from STEM Images of Xanthine Dehydrogenase and Insulin Receptor: Refinements and Molecular Modelling.

Published online by Cambridge University Press:  02 July 2020

D.R. Beniac ,
R.Z.T. Luo ,
A.B. Fernandes ,
T. Iwasaki ,
C.C. Yip  and
F.P. Ottensmeyer
D.R. Beniac
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9, Canada;
R.Z.T. Luo
Affiliation:
Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, 113, Japan.
A.B. Fernandes
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9, Canada;
T. Iwasaki
Affiliation:
Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, M5G 1L6, Canada.
C.C. Yip
Affiliation:
Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, 113, Japan.
F.P. Ottensmeyer
Affiliation:
Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9, Canada;
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Extract

We have reconstructed the three-dimensional quaternary structure of the complete 480 kDa insulin receptor (IR), complexed with Nanogold - labeled insulin, and the 290 kDa Xanthine Dehydrogenase (XDH). Both molecules were imaged by low-dose, low-temperature dark field scanning transmission electron microscopy (STEM). XDH and IR were both freeze-plunged in liquid ethane, and transferred to the STEM (VG HB 601) where they were freeze dried at -130°C. Reconstruction was carried out using the method of quaternion-assisted angular reconstruction (IQAD) as described previously. XDH was further refined by an iterative process in which the IQAD produced volume was used as a reference for further refinements. Separate reconstructions of two sets of half the images indicated an inter-reconstruction resolution of 20 Å and 9 Å by phase residual criteria for the IR and XDH reconstructions, respectively.

Type
Electron Cryomicroscopy of Macromolecules
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 276 - 277
Copyright
Copyright © Microscopy Society of America

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References

1Luo, R.Z-T. et al. Science 285 (1999) 1077.CrossRefGoogle Scholar
2Farrow, N.A. and Ottensmeyer, F.P.J. Opt. Soc. Am. A9 (1992) 1749.CrossRefGoogle Scholar
3Farrow, N.A. and Ottensmeyer, F.P.Ultramicroscopy 52 (1993) 141.CrossRefGoogle Scholar
4Ulrich, A. and Schlessinger, J.Cell 61 (1990) 203.CrossRefGoogle Scholar
5Hille, R.Chem. Rev. 96 (1996) 2757.CrossRefGoogle Scholar
5Dobbek, et al, PNAS 96 (1999) 8884.CrossRefGoogle Scholar
7Hubbard, et al. Nature 372 (1994) 746.CrossRefGoogle Scholar
8Garrett, et al. Nature 394 (1998) 395.CrossRefGoogle Scholar
9Leahy, et al. Proteins. Struct. Funct. 19 (1994) 48.CrossRefGoogle Scholar