scholarly journals A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy

2021 ◽  
Vol 7 (2) ◽  
pp. 118
Author(s):  
Natalia Voskoboynikova ◽  
Maria Karlova ◽  
Rainer Kurre ◽  
Armen Y. Mulkidjanian ◽  
Konstantin V. Shaitan ◽  
...  

The cell wall sensor Wsc1 belongs to a small family of transmembrane proteins, which are crucial to sustain cell integrity in yeast and other fungi. Wsc1 acts as a mechanosensor of the cell wall integrity (CWI) signal transduction pathway which responds to external stresses. Here we report on the purification of Wsc1 by its trapping in water-soluble polymer-stabilized lipid nanoparticles, obtained with an amphipathic styrene-maleic acid (SMA) copolymer. The latter was employed to transfer tagged sensors from their native yeast membranes into SMA/lipid particles (SMALPs), which allows their purification in a functional state, i.e., avoiding denaturation. The SMALPs composition was characterized by fluorescence correlation spectroscopy, followed by two-dimensional image acquisition from single particle transmission electron microscopy to build a three-dimensional model of the sensor. The latter confirms that Wsc1 consists of a large extracellular domain connected to a smaller intracellular part by a single transmembrane domain, which is embedded within the hydrophobic moiety of the lipid bilayer. The successful extraction of a sensor from the yeast plasma membrane by a detergent-free procedure into a native-like membrane environment provides new prospects for in vitro structural and functional studies of yeast plasma proteins which are likely to be applicable to other fungi, including plant and human pathogens.

1990 ◽  
Vol 110 (4) ◽  
pp. 883-894 ◽  
Author(s):  
R Reichelt ◽  
A Holzenburg ◽  
E L Buhle ◽  
M Jarnik ◽  
A Engel ◽  
...  

Nuclear pore complexes (NPCs) prepared from Xenopus laevis oocyte nuclear envelopes were studied in "intact" form (i.e., unexposed to detergent) and after detergent treatment by a combination of conventional transmission electron microscopy (CTEM) and quantitative scanning transmission electron microscopy (STEM). In correlation-averaged CTEM pictures of negatively stained intact NPCs and of distinct NPC components (i.e., "rings," "spoke" complexes, and "plug-spoke" complexes), several fine structural features arranged with octagonal symmetry about a central axis could reproducibly be identified. STEM micrographs of unstained/freeze-dried intact NPCs as well as of their components yielded comparable but less distinct features. Mass determination by STEM revealed the following molecular masses: intact NPC with plug, 124 +/- 11 MD; intact NPC without plug, 112 +/- 11 MD; heavy ring, 32 +/- 5 MD; light ring, 21 +/- 4 MD; plug-spoke complex, 66 +/- 8 MD; and spoke complex, 52 +/- 3 MD. Based on these combined CTEM and STEM data, a three-dimensional model of the NPC exhibiting eightfold centrosymmetry about an axis perpendicular to the plane of the nuclear envelope but asymmetric along this axis is proposed. This structural polarity of the NPC across the nuclear envelope is in accord with its well-documented functional polarity facilitating mediated nucleocytoplasmic exchange of molecules and particles.


Author(s):  
W. D. Cooper ◽  
C. S. Hartley ◽  
J. J. Hren

Interpretation of electron microscope images of crystalline lattice defects can be greatly aided by computer simulation of theoretical contrast from continuum models of such defects in thin foils. Several computer programs exist at the present time, but none are sufficiently general to permit their use as an aid in the identification of the range of defect types encountered in electron microscopy. This paper presents progress in the development of a more general computer program for this purpose which eliminates a number of restrictions contained in other programs. In particular, the program permits a variety of foil geometries and defect types to be simulated.The conventional approximation of non-interacting columns is employed for evaluation of the two-beam dynamical scattering equations by a piecewise solution of the Howie-Whelan equations.


2011 ◽  
Vol 415-417 ◽  
pp. 617-620 ◽  
Author(s):  
Yan Su ◽  
Ying Yun Lin ◽  
Yu Li Fu ◽  
Fan Qian ◽  
Xiu Pei Yang ◽  
...  

Water-soluble gold nanoparticles (AuNPs) were prepared using 2-mercapto-4-methyl-5- thiazoleacetic acid (MMTA) as a stabilizing agent and sodium borohydride (NaBH4) as a reducing agent. The AuNPs product was analyzed by transmission electron microscopy (TEM), UV-vis absorption spectroscopy and Fourier transform infrared spectroscopy (FTIR). The TEM image shows that the particles were well-dispersed and their average particle size is about 5 nm. The UV-vis absorption and FTIR spectra confirm that the MMTA-AuNPs was stabilized by the carboxylate ions present on the surface of the AuNPs.


2008 ◽  
Vol 47 (1) ◽  
pp. 394-399 ◽  
Author(s):  
Hirotoshi Furusho ◽  
Yumiko Mishima ◽  
Norihiro Kameta ◽  
Mitsutoshi Masuda ◽  
Ichiro Yamashita ◽  
...  

2014 ◽  
Vol 47 (5) ◽  
pp. 1729-1735 ◽  
Author(s):  
Xin Nie ◽  
Yimin Guan ◽  
Dongshan Zhao ◽  
Yu Liu ◽  
Jianian Gui ◽  
...  

The crystallographic orientation relationships (ORs) of precipitated β-Mg2Sn particles in Mg–9.76 wt% Sn alloy aged at 573 K for 5 h, corresponding to its peak hardness, were investigated by advanced transmission electron microscopy (TEM). OR-3 of (110)β//(0001)αand [\overline 111]β//[1\overline 210]αand OR-4 of (110)β//(0001)αand [001]β//[2\overline 1\overline 10]αare the key ORs of β-Mg2Sn particles in the alloy. The proportions of β-Mg2Sn particles exhibiting OR-3 and OR-4 were determined as 75.1 and 24.3%, respectively. Crystallographic factors determined the predominance of OR-3 in the precipitated β-Mg2Sn particles. This mechanism was analyzed by a three-dimensional invariant line model constructed using a transformation matrix in reciprocal space. Models of the interface of precipitated β-Mg2Sn and the α-Mg matrix were constructedviahigh-resolution TEM and atomic resolution high-angle annular dark-field scanning TEM.


BioResources ◽  
2006 ◽  
Vol 1 (2) ◽  
pp. 220-232 ◽  
Author(s):  
H. P. S. Abdul Khalil ◽  
M. Siti Alwani ◽  
A. K. Mohd Omar

The chemical composition, anatomical characteristics, lignin distribution, and cell wall structure of oil palm frond (OPF), coconut (COIR), pine-apple leaf (PALF), and banana stem (BS) fibers were analyzed. The chemical composition of fiber was analyzed according to TAPPI Methods. Light microscopy (LM) and transmission electron microscopy (TEM) were used to observe and determine the cell wall structure and lignin distribution of various agro-waste fibers. The results revealed differences in anatomical characteristics, lignin distributions, and cell wall structure of the different types of fibers investigated. Nevertheless, transmission electron microscopy (TEM) micrographs have confirmed that the well wall structure, in each case, could be described in terms of a classical cell wall structure, consisting of primary (P) and secondary (S 1 , S 2 , and S 3 ) layers.


1987 ◽  
Vol 1 (2) ◽  
pp. 322-329 ◽  
Author(s):  
H. Warshawsky

The purpose of this paper is to review evidence which casts doubt on the interpretation universally applied to hexagonal images seen in sectioned enamel. The evidence is based on two possible models to explain the hexagonal profiles seen in mammalian enamel with transmission electron microscopy. The "hexagonal ribbon" model proposes that hexagonal profiles are true cross-sections of elongated hexagonal ribbons. The "rectangular ribbon" model proposes that hexagonal profiles are caused by three-dimensional segments that are parallelepipeds contained in the Epon section. Since shadow projections of such rectangular segments give angles that are inconsistent with the hexagonal unit cell, a model based on ribbons with rhomboidal cut ends and angles of 60 and 120° is proposed. The "rhomboidal ribbon" model projects shadows with angles that are predicted by the unit cell. It is suggested that segments of such crystallites in section project as opaque hexagons on the imaging plane in routine transmission electron microscopy. Morphological observations on crystallites in sections - together with predictions from the hexagonal, rectangular, and rhomboidal ribbon models - indicate that crystallites in rat incisor enamel are flat ribbons with rhomboidal cross-sectional shape. Hexagonal images in electron micrographs of thin-sectioned enamel can result from rhomboidal-ended, parallelepiped-shaped segments of these crystallites projected and viewed as two-dimensional shadows.


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