scholarly journals Use of G-protein fusions to monitor integral membrane protein–protein interactions in yeast

10.1038/80274 ◽  
2000 ◽  
Vol 18 (10) ◽  
pp. 1075-1079 ◽  
Author(s):  
Kathleen N. Ehrhard ◽  
Jörg J. Jacoby ◽  
Xin-Yuan Fu ◽  
Reinhard Jahn ◽  
Henrik G. Dohlman
Keyword(s):  
Membrane Protein ◽  
G Protein ◽  
Protein Interactions ◽  
Integral Membrane Protein ◽  
Protein Protein Interactions

scholarly journals Modulation of Function, Structure and Clustering of K+ Channels by Lipids: Lessons Learnt from KcsA

2020 ◽  
Vol 21 (7) ◽  
pp. 2554 ◽  
Author(s):  
María Lourdes Renart ◽  
Ana Marcela Giudici ◽  
Clara Díaz-García ◽  
María Luisa Molina ◽  
Andrés Morales ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Experimental System ◽  
Integral Membrane Protein ◽  
Lipid Binding ◽  
Common Element ◽  
Protein Protein Interactions ◽  
Expression And Purification ◽  
Arginine Residues ◽  
Channel Currents ◽  
Specific Lipid

KcsA, a prokaryote tetrameric potassium channel, was the first ion channel ever to be structurally solved at high resolution. This, along with the ease of its expression and purification, made KcsA an experimental system of choice to study structure–function relationships in ion channels. In fact, much of our current understanding on how the different channel families operate arises from earlier KcsA information. Being an integral membrane protein, KcsA is also an excellent model to study how lipid–protein and protein–protein interactions within membranes, modulate its activity and structure. In regard to the later, a variety of equilibrium and non-equilibrium methods have been used in a truly multidisciplinary effort to study the effects of lipids on the KcsA channel. Remarkably, both experimental and “in silico” data point to the relevance of specific lipid binding to two key arginine residues. These residues are at non-annular lipid binding sites on the protein and act as a common element to trigger many of the lipid effects on this channel. Thus, processes as different as the inactivation of channel currents or the assembly of clusters from individual KcsA channels, depend upon such lipid binding.

scholarly journals Mechanism of the G-protein mimetic nanobody binding to a muscarinic G-protein-coupled receptor

2018 ◽  
Vol 115 (12) ◽  
pp. 3036-3041 ◽  
Author(s):  
Yinglong Miao ◽  
J. Andrew McCammon
Keyword(s):  
Protein Binding ◽  
G Protein ◽  
Protein Interactions ◽  
Intracellular Signaling ◽  
Md Simulations ◽  
Binding Pocket ◽  
Signaling Proteins ◽  
Protein Protein Interactions ◽  
Intracellular Loops ◽  
G Protein Coupled

Protein–protein binding is key in cellular signaling processes. Molecular dynamics (MD) simulations of protein–protein binding, however, are challenging due to limited timescales. In particular, binding of the medically important G-protein-coupled receptors (GPCRs) with intracellular signaling proteins has not been simulated with MD to date. Here, we report a successful simulation of the binding of a G-protein mimetic nanobody to the M2 muscarinic GPCR using the robust Gaussian accelerated MD (GaMD) method. Through long-timescale GaMD simulations over 4,500 ns, the nanobody was observed to bind the receptor intracellular G-protein-coupling site, with a minimum rmsd of 2.48 Å in the nanobody core domain compared with the X-ray structure. Binding of the nanobody allosterically closed the orthosteric ligand-binding pocket, being consistent with the recent experimental finding. In the absence of nanobody binding, the receptor orthosteric pocket sampled open and fully open conformations. The GaMD simulations revealed two low-energy intermediate states during nanobody binding to the M2 receptor. The flexible receptor intracellular loops contribute remarkable electrostatic, polar, and hydrophobic residue interactions in recognition and binding of the nanobody. These simulations provided important insights into the mechanism of GPCR–nanobody binding and demonstrated the applicability of GaMD in modeling dynamic protein–protein interactions.

Detecting Membrane Protein-protein Interactions Using the Mammalian Membrane Two-hybrid (MaMTH) Assay

2017 ◽  
Vol 9 (1) ◽  
pp. 38-54 ◽  
Author(s):  
Punit Saraon ◽  
Ingrid Grozavu ◽  
Sang Hyun Lim ◽  
Jamie Snider ◽  
Zhong Yao ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Two Hybrid

scholarly journals The mechanism of sperm–oocyte fusion in mammals

Reproduction ◽  
2004 ◽  
Vol 127 (4) ◽  
pp. 423-429 ◽  
Author(s):  
Keisuke Kaji ◽  
Akira Kudo
Keyword(s):  
Membrane Protein ◽  
Sexual Reproduction ◽  
Gene Targeting ◽  
Molecular Mechanism ◽  
Protein Interactions ◽  
Recent Progress ◽  
Research Field ◽  
Protein Protein Interactions ◽  
Long Time ◽  
Adams Family

Sperm–oocyte fusion is one of the most impressive events in sexual reproduction, and the elucidation of its molecular mechanism has fascinated researchers for a long time. Because of the limitation of materials and difficulties in analyzing membrane protein–protein interactions, many attempts have failed to reach this goal. Recent studies involving gene targeting have clearly demonstrated the various molecules that are involved in sperm–oocyte binding and fusion. Sperm ADAMs (family of proteins with a disintegrin and metalloprotease domain), including fertilin α, fertilin β and cyritestin, have been investigated and found to be important for binding rather than for fusion and painstaking studies have raised suspicions that their putative receptors, oocyte integrins, are necessary for the sperm–oocyte interaction. Recently, several studies have focused the spotlight on CD9 and glycosylphosphatidylinositol (GPI)-anchored proteins on oocytes, and epididymal protein DE on sperm, as candidate molecules involved in sperm–oocyte fusion. Lack of, or interference with the function of, these proteins can disrupt the sperm–oocyte fusion without changing the binding. In this review we highlight the candidate molecules involved in the sperm–oocyte interaction suggested from the recent progress in this research field.

A proximity-tagging system to identify membrane protein–protein interactions

2018 ◽  
Vol 15 (9) ◽  
pp. 715-722 ◽  
Author(s):  
Qiang Liu ◽  
Jun Zheng ◽  
Weiping Sun ◽  
Yinbo Huo ◽  
Liye Zhang ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Tagging System
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