Lighting up multiprotein complexes: lessons from GPCR oligomerization

Ciruela, Francisco; Vilardaga, Jean-Pierre; Fernández-Dueñas, Víctor

doi:10.1016/j.tibtech.2010.05.002

Cited by 84 publications

(64 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At higher molecular weights, corresponding to multiples of the monomer, dimeric (∼90-95 kDa) and oligomeric (∼125-135 kDa) receptor species were observed. According to the literature many GPCRs, besides the monomeric form, are expressed as dimers and oligomers, but it is also common for GPCR to appear as homo-and hetero-dimers and oligomers [5,41,33]. Whether APJ multimeric species observed in the fetal and adult pulmonary tissues in our study correspond to homo-or hetero-multiples of the receptor is still not known and could only be clarified by further studies.…”

Section: Discussionmentioning

confidence: 72%

The apelinergic system in the developing lung: Expression and signaling

et al. 2011

View full text Add to dashboard Cite

a b s t r a c tApelin and its receptor APJ constitute a signaling pathway best recognized as an important regulator of cardiovascular homeostasis. This multifunctional peptidergic system is currently being described to be involved in embryonic events which extend into vascular, ocular and heart development. Additionally, it is highly expressed in pulmonary tissue. Therefore, the aim of this study was to investigate the role of apelinergic system during fetal lung development. Immunohistochemistry and Western blot analysis were used to characterize apelin and APJ expression levels and cellular localization in normal fetal rat lungs, at five different gestational ages as well as in the adult. Fetal rat lung explants were cultured in vitro with increasing doses of apelin. Treated lung explants were morphometrically analyzed and assessed for MAPK signaling modifications. Both components of the apelinergic system are constitutively expressed in the developing lung, with APJ exhibiting monomeric, dimeric and oligomeric forms in the pulmonary tissue. Pulmonary epithelium also displayed constitutive nuclear localization of the receptor. Fetal apelin expression is higher than adult expression. Apelin supplementation inhibitory effect on branching morphogenesis was associated with a dose dependent decrease in p38 and JNK phosphorylation. The results presented provide the first evidence of the presence of an apelinergic system operating in the developing lung. Our findings also suggest that apelin inhibits fetal lung growth by suppressing p38 and JNK signaling pathways.

show abstract

Section: Discussionmentioning

confidence: 72%

The apelinergic system in the developing lung: Expression and signaling

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Acceptor photobleaching FRET offers the unique opportunity to investigate protein-protein interactions at a defined subcellular location within the membrane environment of intact cells (5). Analyses were performed following carefully established criteria and involved a large number of measurements for each protein combination.…”

Section: Discussionmentioning

confidence: 99%

“…It has been employed successfully to investigate interactions of membrane proteins, e.g., the G-protein-coupled receptors (5) and nodavirus replicase protein A (7). In acceptor photobleaching FRET, photobleaching of the acceptor results in increased donor emission when the distance between the two is Ͻ10 nm, i.e., when the two proteins or protein segments fused to the fluorophores physically interact (5). Thus, acceptor photobleaching FRET offers the unique opportunity to investigate protein-protein interactions at a defined subcellular location within the membrane environment of intact cells.…”

mentioning

confidence: 99%

Amphipathic α-Helix AH2 Is a Major Determinant for the Oligomerization of Hepatitis C Virus Nonstructural Protein 4B

Gouttenoire

Roingeard

Pénin

et al. 2010

J Virol

View full text Add to dashboard Cite

Nonstructural protein 4B (NS4B) is a key organizer of hepatitis C virus (HCV) replication complex formation. It induces a specific membrane rearrangement, designated membranous web, that serves as a scaffold for the HCV replication complex. However, the mechanisms underlying membranous web formation are poorly understood. Based on fluorescence resonance energy transfer (FRET) and confirmatory coimmunoprecipitation analyses, we provide evidence for an oligomerization of NS4B in the membrane environment of intact cells. Several conserved determinants were found to be involved in NS4B oligomerization, through homotypic and heterotypic interactions. N-terminal amphipathic ␣-helix AH2, comprising amino acids 42 to 66, was identified as a major determinant for NS4B oligomerization. Mutations that affected the oligomerization of NS4B disrupted membranous web formation and HCV RNA replication, implying that oligomerization of NS4B is required for the creation of a functional replication complex. These findings enhance our understanding of the functional architecture of the HCV replication complex and may provide new angles for therapeutic intervention. At the same time, they expand the list of positive-strand RNA virus replicase components acting as oligomers.With an estimated 120 to 180 million chronically infected individuals, hepatitis C virus (HCV) is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma worldwide (26). HCV contains a 9.6-kb positive-strand RNA genome encoding a polyprotein precursor that is co-and posttranslationally processed into 10 structural and nonstructural proteins (28,36). Like all positive-strand RNA viruses investigated thus far, HCV replicates its genome in a membraneassociated replication complex composed of viral proteins, replicating RNA, rearranged intracellular membranes, and additional host factors (6,33,36). Nonstructural protein 4B (NS4B) is the least-characterized HCV protein. However, evidence from biochemical, structural, and genetic studies as well as electron microscopy (EM) indicates that NS4B is a key organizer of HCV replication complex formation (reviewed in reference 16). Indeed, one of the best-documented functions of NS4B is to induce the specific membrane rearrangement, designated membranous web, that serves as a scaffold for the viral replication complex (8, 13). However, the mechanisms underlying membranous web formation are poorly understood.NS4B is a 27-kDa integral membrane protein comprising an N-terminal part (amino acids 1 to ϳ69), a central part harboring four predicted transmembrane segments (amino acids ϳ70 to ϳ190), and a C-terminal part (amino acids ϳ191 to 261).The N-terminal part comprises a predicted and a structurally resolved amphipathic ␣-helix, designated AH1 and AH2, respectively. AH2 comprises amino acids 42 to 66 and has been shown to play an important role in HCV RNA replication (14). Intriguingly, it has the potential to traverse the phospholipid bilayer as a transmembrane segment, likely upon oligomerization (14...

show abstract

“…1,E and F), and approaches such as the heteromer identification technology or complemented donor-acceptor resonance energy transfer (CODA-RET) have also been implemented for the investigation of complexes involving three interacting partners (Fig. 1, G and H) (Ciruela et al, 2010;Cottet et al, 2012;Kaczor et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

G Protein–Coupled Receptor Multimers: A Question Still Open Despite the Use of Novel Approaches

Vischer

Castro

2015

Mol Pharmacol

View full text Add to dashboard Cite

Heteromerization of G protein-coupled receptors (GPCRs) can significantly change the functional properties of involved receptors. Various biochemical and biophysical methodologies have been developed in the last two decades to identify and functionally evaluate GPCR heteromers in heterologous cells, with recent approaches focusing on GPCR complex stoichiometry and stability. Yet validation of these observations in native tissues is still lagging behind for the majority of GPCR heteromers. Remarkably, recent studies, particularly some involving advanced fluorescence microscopy techniques, are contributing to our current knowledge of aspects that were not well known until now, such as GPCR complex stoichiometry and stability. In parallel, a growing effort is being applied to move the field forward into native systems. This short review will highlight recent developments to study the stoichiometry and stability of GPCR complexes and methodologies to detect native GPCR dimers.

show abstract

Lighting up multiprotein complexes: lessons from GPCR oligomerization

Cited by 84 publications

References 87 publications

The apelinergic system in the developing lung: Expression and signaling

The apelinergic system in the developing lung: Expression and signaling

Amphipathic α-Helix AH2 Is a Major Determinant for the Oligomerization of Hepatitis C Virus Nonstructural Protein 4B

G Protein–Coupled Receptor Multimers: A Question Still Open Despite the Use of Novel Approaches

Contact Info

Product

Resources

About