Crystallographic and cryogenic electron microscopic structures and enzymatic characterization of sulfur oxygenase reductase from Sulfurisphaera tokodaii

AbstractSulfur oxygenase reductases (SORs) are present in thermophilic and mesophilic archaea and bacteria, and catalyze oxygen-dependent oxygenation and disproportionation of elemental sulfur. SOR has a hollow, spherical homo-24-mer structure and reactions take place at active sites inside the chamber. The crystal structures of SORs from two Acidianus species have been reported. However, the states of the active site components (mononuclear iron and cysteines) and the entry and exit paths of the substrate and products are still in dispute. Here, we report the biochemical and structural characterizations of SORs from the thermoacidophilic archaeon Sulfurisphaera tokodaii (StSOR) and present high-resolution structures determined by X-ray crystallography and cryogenic electron microscopy (cryo-EM). The crystal structure of StSOR was determined at 1.73 Å resolution. At the catalytic center, iron is ligated to His86, His90, Glu114, and two water molecules. Three conserved cysteines in the cavity are located 9.5∼13 Å from the iron and were observed as free thiol forms. A mutational analysis indicated that the iron and one of the cysteines (Cys31) were essential for both activities and the other two cysteines (Cys101 and Cys104) had a supportive role. The cryo-EM structure was determined at 2.24 Å resolution using an instrument operating at 200 kV. The two structures determined by different methodologies showed similar main chain traces, but the maps exhibited different features at catalytically important components. Given the high resolution achieved in this study, StSOR was shown to be a good benchmark sample for cryo-EM measurements.HighlightsSulfur oxygenase reductase (SOR) was biochemically and structurally characterized.High resolution structures of SOR were determined by crystallography and cryo-EM.Twenty-four identical subunits of SOR form a hollow sphere.Catalytic components exhibited different features in the crystal and cryo-EM structures.

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Electron Microscopic Detection and Characterization of Nonhuman Primate Enteric Viruses

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100054145 ◽

1982 ◽

Vol 40 ◽

pp. 306-307

Author(s):

G. C. Smith ◽

R. L. Heberling ◽

S. S. Kalter

Keyword(s):

Electron Microscopy ◽

Animal Model ◽

Nonhuman Primate ◽

Clinical Condition ◽

Intestinal Tract ◽

Enteric Viruses ◽

Electron Microscopic ◽

Microscopic Detection

A number of viral agents are recognized as and suspected of causing the clinical condition “gastroenteritis.” In our attempts to establish an animal model for studies of this entity, we have been examining the nonhuman primate to ascertain what viruses may be found in the intestinal tract of “normal” animals as well as animals with diarrhea. Several virus types including coronavirus, adenovirus, herpesvirus, and picornavirus (Table I) were detected in our colony; however, rotavirus, astrovirus, and calicivirus have not yet been observed. Fecal specimens were prepared for electron microscopy by procedures reported previously.

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The Immuno-Electron Microscopic Localization of the Mo-Fe Protein Component of Nitrogenase in Cells of Azotobacter Vinelandii

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100073313 ◽

1973 ◽

Vol 31 ◽

pp. 574-575

Author(s):

J. T. Stasny ◽

R. C. Burns ◽

R. W. F. Hardy

Keyword(s):

Cellular Localization ◽

Direct Evidence ◽

Azotobacter Vinelandii ◽

Intracellular Localization ◽

Protein Component ◽

Electron Microscopic ◽

Thin Sections ◽

Fe Protein ◽

Intracytoplasmic Membranes

Structure-functlon studies of biological N2-fixation have correlated the presence of the enzyme nitrogenase with increased numbers of intracytoplasmic membranes in Azotobacter. However no direct evidence has been provided for the internal cellular localization of any nitrogenase. Recent advances concerned with the crystallizatiorTand the electron microscopic characterization of the Mo-Fe protein component of Azotobacter nitrogenase, prompted the use of this purified protein to obtain antibodies (Ab) to be conjugated to electron dense markers for the intracellular localization of the protein by electron microscopy. The present study describes the use of ferritin conjugated to goat antitMo-Fe protein immunoglobulin (IgG) and the observations following its topical application to thin sections of N2-grown Azotobacter.

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High-resolution transmission electron microscopic study of highly oxygen doped silicon layer

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100155438 ◽

1989 ◽

Vol 47 ◽

pp. 692-693

Author(s):

H. Takaoka ◽

M. Tomita ◽

T. Hayashi

Keyword(s):

High Resolution ◽

Oxygen Concentration ◽

Silicon Layer ◽

Detailed Structure ◽

Electron Microscopic ◽

Cross Sectional ◽

Transmission Electron ◽

Doped Silicon ◽

Argon Ion

High resolution transmission electron microscopy (HRTEM) is the effective technique for characterization of detailed structure of semiconductor materials. Oxygen is one of the important impurities in semiconductors. Detailed structure of highly oxygen doped silicon has not clearly investigated yet. This report describes detailed structure of highly oxygen doped silicon observed by HRTEM. Both samples prepared by Molecular beam epitaxy (MBE) and ion implantation were observed to investigate effects of oxygen concentration and doping methods to the crystal structure.The observed oxygen doped samples were prepared by MBE method in oxygen environment on (111) substrates. Oxygen concentration was about 1021 atoms/cm3. Another sample was silicon of (100) orientation implanted with oxygen ions at an energy of 180 keV. Oxygen concentration of this sample was about 1020 atoms/cm3 Cross-sectional specimens of (011) orientation were prepared by argon ion thinning and were observed by TEM at an accelerating voltage of 400 kV.

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Electron microscopic characterization of diamond thin films and substrates for diamond heteroepitaxial growth

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100154937 ◽

1989 ◽

Vol 47 ◽

pp. 592-593

Author(s):

J.B. Posthill ◽

R.P. Burns ◽

R.A. Rudder ◽

Y.H. Lee ◽

R.J. Markunas ◽

...

Keyword(s):

Thin Films ◽

Wide Band ◽

Deposition Process ◽

Heteroepitaxial Growth ◽

Electron Microscopic ◽

Wide Band Gap ◽

Lattice Matching ◽

Different Types ◽

Diamond Thin Films

Because of diamond’s wide band gap, high thermal conductivity, high breakdown voltage and high radiation resistance, there is a growing interest in developing diamond-based devices for several new and demanding electronic applications. In developing this technology, there are several new challenges to be overcome. Much of our effort has been directed at developing a diamond deposition process that will permit controlled, epitaxial growth. Also, because of cost and size considerations, it is mandatory that a non-native substrate be developed for heteroepitaxial nucleation and growth of diamond thin films. To this end, we are currently investigating the use of Ni single crystals on which different types of epitaxial metals are grown by molecular beam epitaxy (MBE) for lattice matching to diamond as well as surface chemistry modification. This contribution reports briefly on our microscopic observations that are integral to these endeavors.

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