Investigating Bcl-2 family protein-protein interactions using a high-speed multiplexing confocal FLIM microscope

2019 ◽  
Author(s):  
Nehad Hirmiz ◽  
Anthony Tsikouras ◽  
Elizabeth Osterlund ◽  
Morgan Richards ◽  
Jessica Kun ◽  
...  
Keyword(s):  
Protein Interactions ◽  
High Speed ◽  
Protein Protein Interactions ◽  
Family Protein

scholarly journals The Role of the Acidity of N-Heteroaryl Sulfonamides as Inhibitors of Bcl-2 Family Protein–Protein Interactions

2013 ◽  
Vol 4 (2) ◽  
pp. 186-190 ◽  
Author(s):  
B. Barry Touré ◽  
Karen Miller-Moslin ◽  
Naeem Yusuff ◽  
Lawrence Perez ◽  
Michael Doré ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Family Protein

scholarly journals Structural titration of receptor ion channel GLIC gating by HS-AFM

2018 ◽  
Vol 115 (41) ◽  
pp. 10333-10338 ◽  
Author(s):  
Yi Ruan ◽  
Kevin Kao ◽  
Solène Lefebvre ◽  
Arin Marchesi ◽  
Pierre-Jean Corringer ◽  
...  
Keyword(s):  
Ion Channel ◽  
Single Molecule ◽  
Conformational Changes ◽  
Protein Interactions ◽  
High Speed ◽  
Conformational Dynamics ◽  
Protein Protein Interactions ◽  
Ion Conductance ◽  
Single Molecule Level ◽  
Selective Channel

Gloeobacter violaceus ligand-gated ion channel (GLIC), a proton-gated, cation-selective channel, is a prokaryotic homolog of the pentameric Cys-loop receptor ligand-gated ion channel family. Despite large changes in ion conductance, small conformational changes were detected in X-ray structures of detergent-solubilized GLIC at pH 4 (active/desensitized state) and pH 7 (closed state). Here, we used high-speed atomic force microscopy (HS-AFM) combined with a buffer exchange system to perform structural titration experiments to visualize GLIC gating at the single-molecule level under native conditions. Reference-free 2D classification revealed channels in multiple conformational states during pH gating. We find changes of protein–protein interactions so far elusive and conformational dynamics much larger than previously assumed. Asymmetric pentamers populate early stages of activation, which provides evidence for an intermediate preactivated state.

scholarly journals Solution Structures of Two CCHC Zinc Fingers from the FOG Family Protein U-Shaped that Mediate Protein–Protein Interactions

Structure ◽  
2000 ◽  
Vol 8 (11) ◽  
pp. 1157-1166 ◽  
Author(s):  
Chu Kong Liew ◽  
Kasper Kowalski ◽  
Archa H Fox ◽  
Anthea Newton ◽  
Belinda K Sharpe ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Zinc Fingers ◽  
Protein Protein Interactions ◽  
Solution Structures ◽  
Family Protein

scholarly journals High-Speed Atomic Force Microscopy of Protein-Protein Interactions

2013 ◽  
Vol 104 (2) ◽  
pp. 386a
Author(s):  
Simon Scheuring ◽  
Ignacio Casuso ◽  
Felix Rico ◽  
Adai Colom
Keyword(s):  
Atomic Force Microscopy ◽  
Protein Interactions ◽  
High Speed ◽  
Protein Protein Interactions ◽  
Force Microscopy ◽  
Atomic Force

scholarly journals Photocatalytic proximity labelling of MCL-1 by a BH3 ligand

2019 ◽  
Vol 2 (1) ◽  
Author(s):  
Hester A. Beard ◽  
Jacob R. Hauser ◽  
Martin Walko ◽  
Rachel M. George ◽  
Andrew J. Wilson ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Visible Light ◽  
Dynamic Analysis ◽  
Protein Interactions ◽  
Cysteine Residue ◽  
Tandem Mass ◽  
Protein Protein Interactions ◽  
Apoptotic Protein ◽  
Family Protein

AbstractLigand-directed protein labelling allows the introduction of diverse chemical functionalities onto proteins without the need for genetically encoded tags. Here we report a method for the rapid labelling of a protein using a ruthenium-bipyridyl (Ru(II)(bpy)3)-modified peptide designed to mimic an interacting BH3 ligand within a BCL-2 family protein-protein interactions. Using sub-stoichiometric quantities of (Ru(II)(bpy)3)-modified NOXA-B and irradiation with visible light for 1 min, the anti-apoptotic protein MCL-1 can be photolabelled with a variety of functional tags. In contrast with previous reports on Ru(II)(bpy)3-mediated photolabelling, tandem mass spectrometry experiments reveal that the labelling site is a cysteine residue of MCL-1. MCL-1 can be labelled selectively in mixtures with other proteins, including the structurally related BCL-2 member, BCL-xL. These results demonstrate that proximity-induced photolabelling is applicable to interfaces that mediate protein-protein interactions, and pave the way towards future use of ligand-directed proximity labelling for dynamic analysis of the interactome of BCL-2 family proteins.

scholarly journals In Situ Structure of the Vibrio Polar Flagellum Reveals a Distinct Outer Membrane Complex and Its Specific Interaction with the Stator

2019 ◽  
Vol 202 (4) ◽  
Author(s):  
Shiwei Zhu ◽  
Tatsuro Nishikino ◽  
Norihiro Takekawa ◽  
Hiroyuki Terashima ◽  
Seiji Kojima ◽  
...  
Keyword(s):  
High Resolution ◽  
Protein Interactions ◽  
High Speed ◽  
Electron Tomography ◽  
Specific Interaction ◽  
Protein Protein Interactions ◽  
Content Type ◽  
Cryo Electron Tomography ◽  
Flagellar Rotation

ABSTRACT The bacterial flagellum is a biological nanomachine that rotates to allow bacteria to swim. For flagellar rotation, torque is generated by interactions between a rotor and a stator. The stator, which is composed of MotA and MotB subunit proteins in the membrane, is thought to bind to the peptidoglycan (PG) layer, which anchors the stator around the rotor. Detailed information on the stator and its interactions with the rotor remains unclear. Here, we deployed cryo-electron tomography and genetic analysis to characterize in situ structure of the bacterial flagellar motor in Vibrio alginolyticus, which is best known for its polar sheathed flagellum and high-speed rotation. We determined in situ structure of the motor at unprecedented resolution and revealed the unique protein-protein interactions among Vibrio-specific features, namely the H ring and T ring. Specifically, the H ring is composed of 26 copies of FlgT and FlgO, and the T ring consists of 26 copies of a MotX-MotY heterodimer. We revealed for the first time a specific interaction between the T ring and the stator PomB subunit, providing direct evidence that the stator unit undergoes a large conformational change from a compact form to an extended form. The T ring facilitates the recruitment of the extended stator units for the high-speed motility in Vibrio species. IMPORTANCE The torque of flagellar rotation is generated by interactions between a rotor and a stator; however, detailed structural information is lacking. Here, we utilized cryo-electron tomography and advanced imaging analysis to obtain a high-resolution in situ flagellar basal body structure in Vibrio alginolyticus, which is a Gram-negative marine bacterium. Our high-resolution motor structure not only revealed detailed protein-protein interactions among unique Vibrio-specific features, the T ring and H ring, but also provided the first structural evidence that the T ring interacts directly with the periplasmic domain of the stator. Docking atomic structures of key components into the in situ motor map allowed us to visualize the pseudoatomic architecture of the polar sheathed flagellum in Vibrio spp. and provides novel insight into its assembly and function.

Protein-protein interactions of the p53 family with the Bcl-2 family in hepatocellular carcinoma

2011 ◽  
Vol 49 (08) ◽  
Author(s):  
LC König ◽  
M Meinhard ◽  
C Sandig ◽  
MH Bender ◽  
A Lovas ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
P53 Family
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