细胞技术

Many transmembrane receptors are regulated by associations with scaffold proteins containing PDZ domains, which interact with receptor carboxyl-termini to control receptor signaling, trafficking, and localization. We describe here approaches for detecting and charac ...

Heterotrimeric G proteins are the main signal-transducing molecules activated by G protein-coupled receptors. Their GTP-dependent activation leads to the regulation of different effectors such as adenylyl cyclases, phospholipases, and RhoGEFs. To understand the full biol ...

β-arrestins, through their scaffolding functions, are key regulators of G protein-coupled receptor (GPCR) signaling and intracellular trafficking. However, little is known about the dynamics of β-arrestin/receptor interactions and how these complexes, and complexes with ot ...

Protein–protein interaction is a widely existing phenomenon and is essential for almost all biological processes, extending from the formation of cellular macromolecular structures and enzymatic complexes to the regulation of signal transduction pathways. Proteins inte ...

G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) comprise a diverse class of signaling molecules involved in cellular physiology and pathology. In recent years, intracellular biosensors have been developed to monitor changes in intracellular cAMP in real time, ...

Advances in imaging assays based on resonance energy transfer (RET) have made it possible to study protein/protein interactions in living cells under physiological conditions. It is now possible to measure the kinetics of changes in these interactions in response to ligand stimulati ...

Cells co-express multiple G protein β and γ subunit isoforms, but the extent to which individual subunits associate to form particular βγ complexes is not known. This issue is important because in vivo knockout experiments suggest that specific βγ complexes may have unique functions despi ...

Over a period of 15 years the concept of G protein-coupled receptor (GPCR) dimerization moved from a challenging hypothesis to a scientific fact, which is now accepted by the vast majority of the scientists working in the field. However, several important issues remain debated such as the biolog ...

Reconstituted high-density lipoprotein particles (rHDL) are powerful platforms used as a model phospholipid bilayer system to study membrane proteins. They consist of a discoidal-shaped planar bilayer of phospholipids that is surrounded by a dimer of apolipoprotein A-I (apoA-I). ...

Voltage-gated calcium channels are key regulators of calcium homeostasis in excitable cells. A number of cellular signaling pathways serve to fine tune calcium channel activity, including the activation of G protein-coupled receptors. Besides regulating channel activity via s ...

G protein-coupled receptors (GPCRs) are key players in cell–cell communication, the dysregulation of which has often deleterious effects leading to pathologies such as psychiatric and neurological diseases. Consequently, GPCRs represent excellent drug targets, and as such are ...

Bioluminescence energy transfer (BRET) has become a powerful tool to study protein–protein interactions and conformational changes among interacting proteins. In particular, BRET assays performed in living cells have revealed that heptahelical receptors (7TMRs), hetero ...

Drug “ligands” that bind G protein-coupled receptors (GPCRs) can either stimulate, fully (full agonists) or partially (partial agonists), or reduce (inverse agonists) basal receptor activity, by stabilizing different receptor conformations. The term “intrinsic efficacy” was ...

The growth and development in the last decade of accurate and reliable mass data collection techniques has greatly enhanced our comprehension of cell signaling networks and pathways. At the same time however, these technological advances have also increased the difficulty of satisfa ...

The Regulator of G protein Signaling (RGS) proteins were identified as a family in 1996 and humans have more than 30 such proteins. Their best known function is to suppress G Protein-Coupled Receptors (GPCR) signaling by increasing the rate of Gα turnoff through stimulation of GTPase activity (i ...

Despite rapid growth in our knowledge of potential disease targets following completion of the first drafts of the human genome over 10 years ago, the success rate of new therapeutic discovery has been frustratingly low. In addition to the widely reported costs and single-digit success rate of ...

In the last 10 years, imaging assays based on resonance energy transfer (RET) and protein fragment complementation have made it possible to study interactions between components of G protein-coupled receptor (GPCR) signalling complexes in living cells under physiological condit ...

The partitioning of secretory and membrane protein-encoding mRNAs to the endoplasmic reticulum (ER), and their translation on ER-associated ribosomes, governs access to the secretory/exocytic pathways of the cell. As mRNAs encoding secretory and membrane proteins comprise ap ...

Isolating mitochondria by subcellular fractionation is a well-established method for retrieving intact and functional mitochondria. This procedure has been used to identify proteins of the mitochondria and to explore import mechanisms. Using the same method, it was shown that mit ...

Asymmetric mRNA localisation to specific compartments of the cell is a fundamental mechanism of �spatial and temporal regulation of gene expression. It is used by a variety of organisms and cell types to achieve different cellular functions. However, the mechanisms of mRNA localisation ...