scholarly journals Streptavidin interfacing as a general strategy to localize fluorescent membrane tension probes in cells

2019 ◽  
Vol 10 (1) ◽  
pp. 310-319 ◽  
Author(s):  
Antoine Goujon ◽  
Karolína Straková ◽  
Naomi Sakai ◽  
Stefan Matile
Keyword(s):  
General Strategy ◽  
Membrane Tension ◽  
Site Specific ◽  
In Cells

Site-specific labeling with biotinylated mechanophores is probed to address the next challenge toward the imaging of forces in cells.

Repurposing Erythrocytes as a “Photoactivatable Bomb”: A General Strategy for Site‐Specific Drug Release in Blood Vessels

Small ◽  
2021 ◽  
pp. 2100753
Author(s):  
Ya‐Xuan Zhu ◽  
Hao‐Ran Jia ◽  
Yuxin Guo ◽  
Xiaoyang Liu ◽  
Ningxuan Zhou ◽  
...  
Keyword(s):  
Drug Release ◽  
Blood Vessels ◽  
General Strategy ◽  
Specific Drug ◽  
Site Specific

scholarly journals Quantitative Chemoproteomics for Site-Specific Analysis of Protein Alkylation by 4-Hydroxy-2-Nonenal in Cells

2015 ◽  
Vol 87 (5) ◽  
pp. 2535-2541 ◽  
Author(s):  
Jing Yang ◽  
Keri A. Tallman ◽  
Ned A. Porter ◽  
Daniel C. Liebler
Keyword(s):  
Site Specific ◽  
Specific Analysis ◽  
In Cells

scholarly journals A general strategy exploiting m5C duplex-remodelling effect for selective detection of RNA and DNA m5C methyltransferase activity in cells

2019 ◽  
Author(s):  
Tianming Yang ◽  
Joanne J A Low ◽  
Esther C Y Woon
Keyword(s):  
Dna Methyltransferase ◽  
General Strategy ◽  
Dna Duplexes ◽  
Methyltransferase Activity ◽  
Cpg Dna ◽  
Sugar Pucker ◽  
High Profile ◽  
New Strategy ◽  
Rna And Dna ◽  
In Cells

Abstract RNA:5-methylcytosine (m5C) methyltransferases are currently the focus of intense research following a series of high-profile reports documenting their physiological links to several diseases. However, no methods exist which permit the specific analysis of RNA:m5C methyltransferases in cells. Herein, we described how a combination of biophysical studies led us to identify distinct duplex-remodelling effects of m5C on RNA and DNA duplexes. Specifically, m5C induces a C3′-endo to C2′-endo sugar-pucker switch in CpG RNA duplex but triggers a B-to-Z transformation in CpG DNA duplex. Inspired by these different ‘structural signatures’, we developed a m5C-sensitive probe which fluoresces spontaneously in response to m5C-induced sugar-pucker switch, hence useful for sensing RNA:m5C methyltransferase activity. Through the use of this probe, we achieved real-time imaging and flow cytometry analysis of NOP2/Sun RNA methyltransferase 2 (NSUN2) activity in HeLa cells. We further applied the probe to the cell-based screening of NSUN2 inhibitors. The developed strategy could also be adapted for the detection of DNA:m5C methyltransferases. This was demonstrated by the development of DNA m5C-probe which permits the screening of DNA methyltransferase 3A inhibitors. To our knowledge, this study represents not only the first examples of m5C-responsive probes, but also a new strategy for discriminating RNA and DNA m5C methyltransferase activity in cells.

scholarly journals Crawling from soft to stiff matrix polarizes the cytoskeleton and phosphoregulates myosin-II heavy chain

2012 ◽  
Vol 199 (4) ◽  
pp. 669-683 ◽  
Author(s):  
Matthew Raab ◽  
Joe Swift ◽  
P.C. Dave P. Dingal ◽  
Palak Shah ◽  
Jae-Won Shin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Myosin Ii ◽  
Stress Fibers ◽  
Site Specific ◽  
Soft Matrix ◽  
3D Collagen ◽  
The Difference ◽  
Rigid Surfaces ◽  
Tissue Matrix ◽  
In Cells

On rigid surfaces, the cytoskeleton of migrating cells is polarized, but tissue matrix is normally soft. We show that nonmuscle MIIB (myosin-IIB) is unpolarized in cells on soft matrix in 2D and also within soft 3D collagen, with rearward polarization of MIIB emerging only as cells migrate from soft to stiff matrix. Durotaxis is the tendency of cells to crawl from soft to stiff matrix, and durotaxis of primary mesenchymal stem cells (MSCs) proved more sensitive to MIIB than to the more abundant and persistently unpolarized nonmuscle MIIA (myosin-IIA). However, MIIA has a key upstream role: in cells on soft matrix, MIIA appeared diffuse and mobile, whereas on stiff matrix, MIIA was strongly assembled in oriented stress fibers that MIIB then polarized. The difference was caused in part by elevated phospho-S1943–MIIA in MSCs on soft matrix, with site-specific mutants revealing the importance of phosphomoderated assembly of MIIA. Polarization is thus shown to be a highly regulated compass for mechanosensitive migration.

A General Strategy for Site-Specific Double Labeling of Globular Proteins for Kinetic FRET Studies

2002 ◽  
Vol 13 (5) ◽  
pp. 1163-1170 ◽  
Author(s):  
V. Ratner ◽  
E. Kahana ◽  
M. Eichler ◽  
E. Haas
Keyword(s):  
Globular Proteins ◽  
General Strategy ◽  
Double Labeling ◽  
Site Specific

scholarly journals Membrane Tension Dictates the Spatiotemporal Heterogeneity of Endocytic Clathrin Coat Dynamics in Cells

2018 ◽  
Vol 114 (3) ◽  
pp. 280a-281a
Author(s):  
Nathan M. Willy ◽  
Joshua Ferguson ◽  
Scott Huber ◽  
Spencer Heidotting ◽  
Esra Aygun ◽  
...  
Keyword(s):  
Membrane Tension ◽  
Spatiotemporal Heterogeneity ◽  
In Cells

scholarly journals Site-Specific Identification of SUMO-2 Targets in Cells Reveals an Inverted SUMOylation Motif and a Hydrophobic Cluster SUMOylation Motif

2010 ◽  
Vol 39 (4) ◽  
pp. 641-652 ◽  
Author(s):  
Ivan Matic ◽  
Joost Schimmel ◽  
Ivo A. Hendriks ◽  
Maria A. van Santen ◽  
Frans van de Rijke ◽  
...  
Keyword(s):  
Site Specific ◽  
Specific Identification ◽  
Hydrophobic Cluster ◽  
In Cells
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