Molecular Packaging of Biocatalysts Using a Robust Protein Cage

10.26434/chemrxiv.12063243 ◽

2020 ◽

Author(s):

Nikola Lončar ◽

Henriette J. Rozeboom ◽

Linda E. Franken ◽

Marc C. A. Stuart ◽

Marco Fraaije

Keyword(s):

Electron Microscopy ◽

Expression System ◽

Size Exclusion ◽

Oligomeric Structure ◽

X Ray Diffraction ◽

X Ray ◽

Protein Cage ◽

Size Exclusion Effect ◽

Exclusion Effect

In this paper, we report on the discovery of a novel, robust protein cage (encapsulin) that we could use for packaging various biocatalysts. We have elucidated the structure of the stable encapsulin by electron microscopy and X-ray diffraction. Furthermore, we developed an effective expression system for the encapsulin and a facile protocol for preparing encapsulated enzymes. By packaging and testing various enzymes (varying in size, oligomeric structure, and cofactor type) we demonstrate that, through encapsulation, the enzymes become significantly more stable. We also provide evidence that the pores of the encapsulin, through a size-exclusion effect, can modulate the substrate acceptance profile of an encapsulated enzyme.

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Toscana virusnucleoprotein oligomer organization observed in solution

Acta Crystallographica Section D Structural Biology ◽

10.1107/s2059798317008774 ◽

2017 ◽

Vol 73 (8) ◽

pp. 650-659 ◽

Cited By ~ 4

Author(s):

Amal Baklouti ◽

Adeline Goulet ◽

Julie Lichière ◽

Bruno Canard ◽

Rémi N. Charrel ◽

...

Keyword(s):

Electron Microscopy ◽

Size Exclusion ◽

Integrative Approach ◽

X Ray Diffraction ◽

X Ray ◽

Transmission Electron ◽

Closed Ring ◽

Rnp Complex ◽

Exclusion Chromatography ◽

Nucleoprotein N

Toscana virus(TOSV) is an arthropod-borne virus belonging to thePhlebovirusgenus within theBunyaviridaefamily. As in other bunyaviruses, the genome of TOSV is made up of three RNA segments. They are encapsidated by the nucleoprotein (N), which also plays an essential role in virus replication. To date, crystallographic structures of phlebovirus N have systematically revealed closed-ring organizations which do not fully match the filamentous organization of the ribonucleoprotein (RNP) complex observed by electron microscopy. In order to further bridge the gap between crystallographic data on N and observations of the RNP by electron microscopy, the structural organization of recombinant TOSV N was investigated by an integrative approach combining X-ray diffraction crystallography, transmission electron microscopy, small-angle X-ray scattering, size-exclusion chromatography and multi-angle laser light scattering. It was found that in solution TOSV N forms open oligomers consistent with the encapsidation mechanism of phlebovirus RNA.

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Electron Microscopy of Human Gallstones

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065523 ◽

1971 ◽

Vol 29 ◽

pp. 296-297

Author(s):

C. Wolpers ◽

R. Blaschke

Keyword(s):

Electron Microscopy ◽

Room Temperature ◽

Bile Pigment ◽

X Ray Diffraction ◽

Triclinic Crystal System ◽

X Ray ◽

Follow Up Study ◽

Infra Red ◽

Main Components

Scanning microscopy was used to study the surface of human gallstones and the surface of fractures. The specimens were obtained by operation, washed with water, dried at room temperature and shadowcasted with carbon and aluminum. Most of the specimens belong to patients from a series of X-ray follow-up study, examined during the last twenty years. So it was possible to evaluate approximately the age of these gallstones and to get information on the intensity of growing and solving.Cholesterol, a group of bile pigment substances and different salts of calcium, are the main components of human gallstones. By X-ray diffraction technique, infra-red spectroscopy and by chemical analysis it was demonstrated that all three components can be found in any gallstone. In the presence of water cholesterol crystallizes in pane-like plates of the triclinic crystal system.

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Electron Microscopy of Shock Loaded Polycrystalline Beryllium

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052547 ◽

1974 ◽

Vol 32 ◽

pp. 506-507 ◽

Cited By ~ 1

Author(s):

J. M. Galbraith ◽

L. E. Murr ◽

A. L. Stevens

Keyword(s):

Electron Microscopy ◽

Wave Propagation ◽

Structural Change ◽

Single Crystal ◽

Diffraction Pattern ◽

Shock Loading ◽

Slip Systems ◽

Compression Tests ◽

X Ray Diffraction ◽

X Ray

Uniaxial compression tests and hydrostatic tests at pressures up to 27 kbars have been performed to determine operating slip systems in single crystal and polycrystal1ine beryllium. A recent study has been made of wave propagation in single crystal beryllium by shock loading to selectively activate various slip systems, and this has been followed by a study of wave propagation and spallation in textured, polycrystal1ine beryllium. An alteration in the X-ray diffraction pattern has been noted after shock loading, but this alteration has not yet been correlated with any structural change occurring during shock loading of polycrystal1ine beryllium.This study is being conducted in an effort to characterize the effects of shock loading on textured, polycrystal1ine beryllium. Samples were fabricated from a billet of Kawecki-Berylco hot pressed HP-10 beryllium.

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Freeze-Fracture Replication of Frozen Outer Segments of Rat Retinal Rods

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100063871 ◽

1969 ◽

Vol 27 ◽

pp. 392-393

Author(s):

Thomas S. Leeson ◽

C. Roland Leeson

Keyword(s):

Electron Microscopy ◽

Cross Section ◽

Outer Segment ◽

Freeze Fracture ◽

X Ray Diffraction ◽

X Ray ◽

Outer Segments ◽

Retinal Rods ◽

Temperature Electron ◽

Freeze Etching

Numerous previous studies of outer segments of retinal receptors have demonstrated a complex internal structure of a series of transversely orientated membranous lamellae, discs, or saccules. In cones, these lamellae probably are invaginations of the covering plasma membrane. In rods, however, they appear to be isolated and separate discs although some authors report interconnections and some continuities with the surface near the base of the outer segment, i.e. toward the inner segment. In some species, variations have been reported, such as longitudinally orientated lamellae and lamellar whorls. In cross section, the discs or saccules show one or more incisures. The saccules probably contain photolabile pigment, with resulting potentials after dipole formation during bleaching of pigment. Continuity between the lamina of rod saccules and extracellular space may be necessary for the detection of dipoles, although such continuity usually is not found by electron microscopy. Particles on the membranes have been found by low angle X-ray diffraction, by low temperature electron microscopy and by freeze-etching techniques.

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A High Resolution Study of G.P. Zones in Aluminum - 4.2 at % Silver

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100055904 ◽

1982 ◽

Vol 40 ◽

pp. 722-723 ◽

Cited By ~ 1

Author(s):

R. Gronsky

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

High Resolution ◽

Computer Simulations ◽

Structural Characteristics ◽

X Ray Diffraction ◽

X Ray ◽

Transmission Electron ◽

Resolution Study

The phenomenon of clustering in Al-Ag alloys has been extensively studied since the early work of Guinierl, wherein the pre-precipitation state was characterized as an assembly of spherical, ordered, silver-rich G.P. zones. Subsequent x-ray and TEM investigations yielded results in general agreement with this model. However, serious discrepancies were later revealed by the detailed x-ray diffraction - based computer simulations of Gragg and Cohen, i.e., the silver-rich clusters were instead octahedral in shape and fully disordered, atleast below 170°C. The object of the present investigation is to examine directly the structural characteristics of G.P. zones in Al-Ag by high resolution transmission electron microscopy.

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Identification of calcium oxalate crystals in dried or fixed tobacco leaf

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111550 ◽

1984 ◽

Vol 42 ◽

pp. 288-289

Author(s):

Vicki L. Baliga ◽

Mary Ellen Counts

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Calcium Oxalate ◽

Growth And Development ◽

Polarized Light ◽

Calcium Oxalate Crystals ◽

X Ray Diffraction ◽

X Ray ◽

Transmission Electron ◽

Scanning Electron

Calcium is an important element in the growth and development of plants and one form of calcium is calcium oxalate. Calcium oxalate has been found in leaf seed, stem material plant tissue culture, fungi and lichen using one or more of the following methods—polarized light microscopy (PLM), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and x-ray diffraction.Two methods are presented here for qualitatively estimating calcium oxalate in dried or fixed tobacco (Nicotiana) leaf from different stalk positions using PLM. SEM, coupled with energy dispersive x-ray spectrometry (EDS), and powder x-ray diffraction were used to verify that the crystals observed in the dried leaf with PLM were calcium oxalate.

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Environmental scanning electron microscopy of fresh human gallstones reveals new morphologies of precipitated calcium salts

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100131243 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1322-1323

Author(s):

Howard S. Kaufman ◽

Keith D. Lillemoe ◽

John T. Mastovich ◽

Henry A. Pitt

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Environmental Scanning Electron Microscopy ◽

Environmental Scanning ◽

X Ray Diffraction ◽

Calcium Salts ◽

Cholesterol Gallstones ◽

X Ray ◽

Ca Carbonate ◽

Scanning Electron

Gallstones contain precipitated cholesterol, calcium salts, and proteins. Calcium (Ca) bilirubinate, palmitate, phosphate, and carbonate occurring in gallstones have variable morphologies but characteristic windowless energy dispersive x-ray (EDX) spectra. Previous studies of gallstone microstructure and composition using scanning electron microscopy (SEM) with EDX have been limited to dehydrated samples. In this state, Ca bilirubinates appear as either glassy masses, which predominate in black pigment stones, or as clusters, which are found mostly in cholesterol gallstones. The three polymorphs of Ca carbonate, calcite, vaterite, and aragonite, have been identified in gallstones by x-ray diffraction, however; the morphologies of these crystals vary in the literature. The purpose of this experiment was to study fresh gallstones by environmental SEM (ESEM) to determine if dehydration affects gallstone Ca salt morphology.Gallstones and bile were obtained fresh at cholecystectomy from 6 patients. To prevent dehydration, stones were stored in bile at 37°C. All samples were studied within 4 days of procurement.

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Electron Microscopy/Diffraction Study of the Ternary Mn-Er-S System

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100179130 ◽

1990 ◽

Vol 48 (1) ◽

pp. 76-77

Author(s):

A. R. Landa Canovas ◽

L.C. Otero Diaz ◽

T. White ◽

B.G. Hyde

Keyword(s):

Electron Microscopy ◽

Gas Flow ◽

Graphite Crucible ◽

Nominal Composition ◽

X Ray Diffraction ◽

Alumina Crucible ◽

X Ray ◽

Intermediate Phases ◽

Twin Band ◽

Ar Gas

X-Ray diffraction revealed two intermediate phases in the system MnS+Er2S3,:MnEr2S4= MnS.Er2S3, and MnEr4S7= MnS.2Er2S3. Their structures may be described as NaCl type, chemically twinned at the unit cell level, and isostructural with CaTi2O4, and Y5S7 respectively; i.e. {l13} NaCl twin band widths are (4,4) and (4,3).The present study was to search for structurally-related (twinned B.) structures and or possible disorder, using the more sensitive and appropiate technigue of electron microscopy/diffraction.A sample with nominal composition MnEr2S4 was made by heating Mn3O4 and Er2O3 in a graphite crucible and a 5% H2S in Ar gas flow at 1500°C for 4 hours. A small amount of this material was thenannealed, in an alumina crucible, contained in sealed evacuated silica tube, for 24 days at 1100°C. Both samples were studied by X-ray powder diffraction, and in JEOL 2000 FX and 4000 EX microscopes.

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