Towards a universal method for protein refolding: The trimeric beta barrel membrane Omp2a as a test case

2012 ◽  
Vol 110 (2) ◽  
pp. 417-423 ◽  
Author(s):  
Guillaume Roussel ◽  
Eric A. Perpète ◽  
André Matagne ◽  
Emmanuel Tinti ◽  
Catherine Michaux
Keyword(s):  
Protein Refolding ◽  
Test Case ◽  
Universal Method ◽  
Beta Barrel

scholarly journals POLArIS, a versatile probe for molecular orientation, revealed actin filaments associated with microtubule asters in early embryos

2021 ◽  
Vol 118 (11) ◽  
pp. e2019071118
Author(s):  
Ayana Sugizaki ◽  
Keisuke Sato ◽  
Kazuyoshi Chiba ◽  
Kenta Saito ◽  
Masahiko Kawagishi ◽  
...  
Keyword(s):  
Actin Filaments ◽  
Molecular Orientation ◽  
Fluorescent Protein ◽  
Three Dimensional ◽  
Super Resolution ◽  
Living Cells ◽  
Test Case ◽  
Polarization Microscopy ◽  
Universal Method ◽  
Early Embryos

Biomolecular assemblies govern the physiology of cells. Their function often depends on the changes in molecular arrangements of constituents, both in the positions and orientations. While recent advancements of fluorescence microscopy including super-resolution microscopy have enabled us to determine the positions of fluorophores with unprecedented accuracy, monitoring the orientation of fluorescently labeled molecules within living cells in real time is challenging. Fluorescence polarization microscopy (FPM) reports the orientation of emission dipoles and is therefore a promising solution. For imaging with FPM, target proteins need labeling with fluorescent probes in a sterically constrained manner, but because of difficulties in the rational three-dimensional design of protein connection, a universal method for constrained tagging with fluorophore was not available. Here, we report POLArIS, a genetically encoded and versatile probe for molecular orientation imaging. Instead of using a direct tagging approach, we used a recombinant binder connected to a fluorescent protein in a sterically constrained manner that can target specific biomolecules of interest by combining with phage display screening. As an initial test case, we developed POLArISact, which specifically binds to F-actin in living cells. We confirmed that the orientation of F-actin can be monitored by observing cells expressing POLArISact with FPM. In living starfish early embryos expressing POLArISact, we found actin filaments radially extending from centrosomes in association with microtubule asters during mitosis. By taking advantage of the genetically encoded nature, POLArIS can be used in a variety of living specimens, including whole bodies of developing embryos and animals, and also be expressed in a cell type/tissue specific manner.

scholarly journals POLArIS, a versatile probe for molecular orientation, revealed actin filaments associated with microtubule asters in early embryos

2020 ◽  
Author(s):  
Ayana Sugizaki ◽  
Keisuke Sato ◽  
Kazuyoshi Chiba ◽  
Kenta Saito ◽  
Masahiko Kawagishi ◽  
...  
Keyword(s):  
Actin Filaments ◽  
Molecular Orientation ◽  
Fluorescent Protein ◽  
Three Dimensional ◽  
Super Resolution ◽  
Living Cells ◽  
Test Case ◽  
Polarization Microscopy ◽  
Universal Method ◽  
Early Embryos

AbstractBiomolecular assemblies govern the physiology of cells. Their function often depends on the changes in molecular arrangements of constituents, both in the positions and orientations. While recent advancements of fluorescence microscopy including super-resolution microscopy have enabled us to determine the positions of fluorophores with unprecedented accuracy, monitoring orientation of fluorescently labeled molecules within living cells in real-time is challenging. Fluorescence polarization microscopy (FPM) reports the orientation of emission dipoles and is therefore a promising solution. For imaging with FPM, target proteins need labeling with fluorescent probes in a sterically constrained manner, but due to difficulties in the rational three-dimensional design of protein connection, universal method for constrained tagging with fluorophore was not available. Here we report POLArIS, a genetically encoded and versatile probe for molecular orientation imaging. Instead of using a direct tagging approach, we used a recombinant binder connected to a fluorescent protein in a sterically constrained manner and can target arbitrary biomolecules by combining with phage-display screening. As an initial test case of POLArIS, we developed POLArISact, which specifically binds to F-actin in living cells. We confirmed that the orientation of F-actin can be monitored by observing cells expressing POLArISact with FPM. In living starfish early embryos expressing POLArISact, we found actin filaments radially extending from centrosomes in association with microtubule asters during mitosis. By taking advantage of the genetically encoded nature, POLArIS can be used in a variety of living specimens including whole bodies of developing embryos and animals, and also expressed in a cell-type/tissue specific manner.

Representation of information in SEM by equal signal lines

1978 ◽  
Vol 36 (1) ◽  
pp. 144-145
Author(s):  
E. Rau ◽  
N. Karelin ◽  
V. Dukov ◽  
M. Kolomeytsev ◽  
S. Gavrikov ◽  
...  
Keyword(s):  
Electronic System ◽  
Space Distribution ◽  
Specimen Surface ◽  
Universal Method ◽  
Base Level ◽  
Signal Value ◽  
Information Signal ◽  
Fixed Base ◽  
Scanning Line ◽  
Amplitude Analyzer

There are different methods and devices for the increase of the videosignal information in SEM. For example, with the help of special pure electronic [1] and opto-electronic [2] systems equipotential areas on the specimen surface in SEM were obtained. This report generalizes quantitative universal method for space distribution representation of research specimen parameter by contour equal signal lines. The method is based on principle of comparison of information signal value with the fixed levels.Transformation image system for obtaining equal signal lines maps was developed in two versions:1)In pure electronic system [3] it is necessary to compare signal U (see Fig.1-a), which gives potential distribution on specimen surface along each scanning line with fixed base level signals εifor obtaining quantitative equipotential information on solid state surface. The amplitude analyzer-comparator gives flare sport videopulses at any fixed coordinate and any instant time when initial signal U is equal to one of the base level signals ε.

Religious Melancholy in the Romantic Period: William Cowper as Test Case

Romanticism ◽  
2009 ◽  
Vol 15 (2) ◽  
pp. 144-155 ◽  
Author(s):  
Jane Darcy
Keyword(s):  
Test Case ◽  
Romantic Period ◽  
William Cowper

From Hard Money to Branch Banking

2016 ◽  
Vol 93 (1) ◽  
pp. 26-44
Author(s):  
Larry Schweikart ◽  
Lynne Pierson Doti
Keyword(s):  
West Coast ◽  
Financial Sector ◽  
Gold Rush ◽  
Test Case ◽  
The West ◽  
Branch Banking

In Gold Rush–era California, banking and the financial sector evolved in often distinctive ways because of the Gold Rush economy. More importantly, the abundance of gold on the West Coast provided an interesting test case for some of the critical economic arguments of the day, especially for those deriving from the descending—but still powerful—positions of the “hard money” Jacksonians.

Biblical Literacy and the English King James Liberal Bible

2014 ◽  
Vol 7 (2) ◽  
pp. 197-211
Author(s):  
James Crossley
Keyword(s):  
Cultural Heritage ◽  
Test Case ◽  
Biblical Literacy ◽  
The Public ◽  
Public Arena ◽  
The Bible ◽  
Liberal Capitalism ◽  
The Rich ◽  
Popular Nationalism ◽  
King James Bible

Using the 400th anniversary of the King James Bible as a test case, this article illustrates some of the important ways in which the Bible is understood and consumed and how it has continued to survive in an age of neoliberalism and postmodernity. It is clear that instant recognition of the Bible-as-artefact, multiple repackaging and pithy biblical phrases, combined with a popular nationalism, provide distinctive strands of this understanding and survival. It is also clear that the KJV is seen as a key part of a proud English cultural heritage and tied in with traditions of democracy and tolerance, despite having next to nothing to do with either. Anything potentially problematic for Western liberal discourse (e.g. calling outsiders “dogs,” smashing babies heads against rocks, Hades-fire for the rich, killing heretics, using the Bible to convert and colonize, etc.) is effectively removed, or even encouraged to be removed, from such discussions of the KJV and the Bible in the public arena. In other words, this is a decaffeinated Bible that has been colonized by, and has adapted to, Western liberal capitalism.

TEST-CASE NO 16: IMPACT OF A DROP ON A THIN FILM OF THE SAME LIQUID (PE, PA)

2004 ◽  
Vol 16 (1-3) ◽  
pp. 105-109
Author(s):  
F. Pigeonneau ◽  
Francois Feuillebois
Keyword(s):  
Thin Film ◽  
Test Case

TEST-CASE NO 15: PHASE INVERSION IN A CLOSED BOX (PC)

2004 ◽  
Vol 16 (1-3) ◽  
pp. 101-104 ◽  
Author(s):  
S. Vincent ◽  
J.-P. Caltagirone ◽  
D. Jamet
Keyword(s):  
Phase Inversion ◽  
Test Case

TEST-CASE NO 7B: ISOTHERMAL VAPORIZATION DUE TO PISTON ASPIRATION (PA)

2004 ◽  
Vol 16 (1-3) ◽  
pp. 61-67
Author(s):  
D. Jamet
Keyword(s):  
Test Case
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