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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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