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Montagne, and T. Meerloo contributed experimental work. Trejo contributed to project design and manuscript preparation. Immunolabeled cells were processed and imaged by confocal microscopy.

The colocalization of PAR1 and LAMP1 is shown in yellow in the merged image and is representative of many cells examined in three independent experiments.

Insets show magnifications of boxed areas. Cells were fixed and processed for immuno-EM. Dual-labeled ultrathin sections revealed that PAR1 WT and 0K mutant detected with nm gold-conjugated secondary antibody black arrows sorted to ILVs present in CDpositive MVBs, which was detected with 6-nm gold-labeled secondary antibody white arrows.

Bar, nm. The asterisk indicates a nonspecific band. Cell lysates were immunoblotted with anti-GFP and -actin antibodies.

The insets are magnifications of the boxed areas. Cell lysates were immunoblotted with anti-HA and -actin antibodies. Cell lysates were immunoblotted with anti-HA and -actin antibodies as controls.

Equivalent amounts of cell lysates were immunoprecipitated IP and examined as described in Fig. IB, immunoblotting. Equivalent amounts of cell lysates were immunoprecipitated and analyzed as described in Fig.

PAR degradation was assessed by immunoblotting IB. Endogenous PAR1 was detected by immunoblotting. Cell lysates were immunoblotted with anti-ALIX and -actin antibodies.

The conserved residues of the YPX 3 L motif are shaded in gray. Cell lysates were immunoprecipitated and examined as described in Fig.

Pulldowns PD were analyzed for the presence of bound protein by immunoblotting. Input was analyzed by Coomassie staining.

PAR1 degradation was analyzed as described in Fig. Membranes were isolated and treated with proteinase K or proteinase K with 0.

Sign In or Create an Account. Advanced Search. User Tools. Sign In. Article Navigation. Article April 30 Dores , Michael R.

This Site. Google Scholar. Buxin Chen , Buxin Chen. Huilan Lin , Huilan Lin. Unice J. Soh , Unice J. May M. Paing , May M.

Louis, MO William A. Montagne , William A. Timo Meerloo , Timo Meerloo. Correspondence to JoAnn Trejo: joanntrejo ucsd. Author and Article Information.

Buxin Chen. Huilan Lin. Timo Meerloo. JoAnn Trejo. Received: October 06 Accepted: March 22 Online Issn: J Cell Biol 3 : — Article history Received:.

Connected Content. Ubiquitin isn't ubiquitous in receptor trafficking. Standard View Views Icon Views. Get Permissions. Cite Icon Cite.

Protease-activated receptor signalling, endocytic sorting and dysregulation in cancer. Search ADS. Persistent signaling by dysregulated thrombin receptor trafficking promotes breast carcinoma cell invasion.

Tsg and the vacuolar protein sorting pathway are essential for HIV-1 budding. Degradation of AP2 during reticulocyte maturation enhances binding of hsc70 and Alix to a common site on TFR for sorting into exosomes.

An essential role for SNX1 in lysosomal sorting of protease-activated receptor Evidence for retromer-, Hrs-, and Tsgindependent functions of sorting nexins.

Regulation of GPCRs by endocytic membrane trafficking and its potential implications. Hepatocyte growth factor-regulated tyrosine kinase substrate HRS mediates post-endocytic trafficking of protease-activated receptor 2 and calcitonin receptor-like receptor.

Role of mammalian vacuolar protein-sorting proteins in endocytic trafficking of a non-ubiquitinated G protein-coupled receptor to lysosomes.

HIV-1 and Ebola virus encode small peptide motifs that recruit Tsg to sites of particle assembly to facilitate egress.

Divergent retroviral late-budding domains recruit vacuolar protein sorting factors by using alternative adaptor proteins.

Direct binding to Rsp5 mediates ubiquitin-independent sorting of Sna3 via the multivesicular body pathway. Bro1 is an endosome-associated protein that functions in the MVB pathway in Saccharomyces cerevisiae.

Characterization of multiple multivesicular body sorting determinants within Sna3: A role for the ubiquitin ligase Rsp5. Clathrin adaptor AP2 regulates thrombin receptor constitutive internalization and endothelial cell resensitization.

Phosphorylation of protease-activated receptor-2 differentially regulates desensitization and internalization. Caveolae are required for protease-selective signaling by protease-activated receptor N-linked glycosylation of protease-activated receptor-1 second extracellular loop: A critical determinant for ligand-induced receptor activation and internalization.

Equine infectious anemia virus utilizes host vesicular protein sorting machinery during particle release. A lumenal domain-dependent pathway for sorting to intralumenal vesicles of multivesicular endosomes involved in organelle morphogenesis.

Ceramide triggers budding of exosome vesicles into multivesicular endosomes. The cytoplasmic tails of protease-activated receptor-1 and substance P receptor specify sorting to lysosomes versus recycling.

Termination of signaling by protease-activated receptor-1 is linked to lysosomal sorting. Protease-activated receptor-1 down-regulation: A mutant HeLa cell line suggests novel requirements for PAR1 phosphorylation and recruitment to clathrin-coated pits.

Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Direct binding to Rsp5p regulates ubiquitination-independent vacuolar transport of Sna3p.

Ubiquitination differentially regulates clathrin-dependent internalization of protease-activated receptor Abbreviations used in this paper:.

Author notes M. Chen contributed equally to this paper. Supplementary data Supplemental material PDF - pdf file.

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