Comparative proteomics of clathrin-coated vesicles

Borner, Georg H. H.; Harbour, Michael E.; Hester, Svenja; Lilley, Kathryn S.; Robinson, Margaret S.

doi:10.1083/jcb.200607164

Cited by 151 publications

(176 citation statements)

References 26 publications

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“…We next investigated whether SERT traversed through Rab4-containing endosomes (34,36). CHO transfectants expressing YFP-Rab4 constructs in combination with CFP-SERT were pretreated with 0, 10, or 100 M 5HT for 40 min and imaged using confocal microscopy.…”

Section: Sert Expression On Cell Membranementioning

confidence: 99%

Serotonin Transamidates Rab4 and Facilitates Its Binding to the C Terminus of Serotonin Transporter

Ahmed

Jeffus

Bukhari

et al. 2008

Journal of Biological Chemistry

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The serotonin transporter (SERT) on the plasma membrane is the major mechanism for the clearance of plasma serotonin (5-hydroxytryptamine (5HT)). The uptake rates of cells depend on the density of SERT molecules on the plasma membrane. Interestingly, the number of SERT molecules on the platelet surface is down-regulated when plasma 5HT ([5HT] ex ) is elevated. It is well reported that stimulation of cells with high [5HT] ex induces transamidation of a small GTPase, Rab4. Modification with 5HT stabilizes Rab4 in its active, GTP-bound form, Rab4-GTP. Although investigating the mechanism by which elevated plasma 5HT level down-regulates the density of SERT molecules on the plasma membrane, we studied Rab4 and SERT in heterologous and platelet expression systems. Our data demonstrate that, in response to elevated [5HT] ex , Rab4-GTP co-localizes with and binds to SERT. The association of SERT with Rab4-GTP depends on: (i) 5HT modification and (ii) the GTP-binding ability of Rab4. Their association retains transporter molecules intracellularly. Furthermore, we mapped the Rab4-SERT association domain to amino acids 616 -624 in the cytoplasmic tail of SERT. This finding provides an explanation for the role of the C terminus in the localization and trafficking of SERT via Rab4 in a plasma 5HT-dependent manner. Therefore, we propose that elevated [5HT] ex "paralyzes" the translocation of SERT from intracellular locations to the plasma membrane by controlling transamidation and Rab4-GTP formation.The serotonin transporter (SERT) 2 is a member of the Cl Ϫ -and Na ϩ -dependent monoamine transporter family, which also includes the dopamine transporter (DAT) and the norepinephrine transporter. SERT is a 630-amino acid plasma membrane-bound glycoprotein. Hydropathy analysis predicts that SERT contains 12 transmembrane domains and that both the N and C termini are exposed to the cytoplasm. The primary function of SERT in the central nervous system involves the regulation of serotonergic signaling via transport of serotonin (5-hydroxytryptamine (5HT)) molecules from the synaptic cleft into the pre-synaptic terminal for re-utilization. SERT is also expressed in non-neuronal cells, including platelets, placental, intestinal and adrenal cell lines, but the exact function of SERT in these cell lines is still under investigation (1-5).The C-and N-terminal regions of monoamine transporter proteins have just recently garnered increased attention for their importance in transport function and localization. Significant work has been accomplished in identifying the importance of the C-terminal region of DAT and norepinephrine transporter in transporter function, expression, and localization (6 -9).The proteins interacting with the N terminus of SERT are syntaxin 1A (10, 11) and secretory carrier membrane protein 2 (12). SERT also complexes with Hic-5 (13) and ␣-synuclein (14), but the functional significance of these interactions are not known. PICK1 (15), MacMARCKS (16), the actin cytoskeleton (49), neuronal nitric-oxide synthase, and Se...

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Section: Sert Expression On Cell Membranementioning

confidence: 99%

Serotonin Transamidates Rab4 and Facilitates Its Binding to the C Terminus of Serotonin Transporter

Ahmed

Jeffus

Bukhari

et al. 2008

Journal of Biological Chemistry

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“…The principal components of coated vesicles isolated from various sources are the heavy and light chains of clathrin and the four subunits of the heterotetrameric "adaptor complexes" (Kirchhausen 2000;Blondeau et al 2004;Girard et al 2005;Borner et al 2006Borner et al , 2012. The heterotetrameric adaptors link the clathrin coat and the membrane bilayer, and they are the principal cargo-recognition molecules.…”

Section: Structure and Biochemistry Of Clathrin-coated Vesiclesmentioning

confidence: 99%

Molecular Structure, Function, and Dynamics of Clathrin-Mediated Membrane Traffic

Kirchhausen

Owen

Harrison

2014

Cold Spring Harbor Perspectives in Biology

411

437

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Clathrin is a molecular scaffold for vesicular uptake of cargo at the plasma membrane, where its assembly into cage-like lattices underlies the clathrin-coated pits of classical endocytosis. This review describes the structures of clathrin, major cargo adaptors, and other proteins that participate in forming a clathrin-coated pit, loading its contents, pinching off the membrane as a lattice-enclosed vesicle, and recycling the components. It integrates as much of the structural information as possible at the time of writing into a sketch of the principal steps in coated-pit and coated-vesicle formation.

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“…The membrane was blocked with 0.25% glycine, 0.1% Tween-20 in PBS, followed by 20% nonfat skimmed milk, then incubated with antibodies diluted in 2% milk, 0.5% Tween-20 in PBS. SK1775 rabbit anti-Sdmg1 antibodies (Best et al, 2008) were used at 0.25 g/ml for Western blotting, rabbit anti-Vamp8 antibodies (Borner et al, 2006) were used at a 1:200 dilution, and mouse anti-Gapdh antibodies (Abcam) were used at a 1:1,000 dilution. Peroxidase-linked secondary antibodies were detected using enhanced chemiluminescence.…”

Section: Western Blottingmentioning

confidence: 99%

Sdmg1 is a component of secretory granules in mouse secretory exocrine tissues

Best

Adams

2008

Developmental Dynamics

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Sdmg1 is a conserved eukaryotic transmembrane protein that is mainly expressed in the gonads where it may have a role in mediating signaling between somatic cells and germ cells. In this study we demonstrate that secretory exocrine cells in the pancreas, salivary gland, and mammary gland also express Sdmg1. Furthermore, we show that Sdmg1 expression is up-regulated during pancreas development when regulated secretory granules start to appear, and that Sdmg1 colocalizes with secretory granule markers in adult pancreatic acinar cells. In addition, we show that Sdmg1 co-purifies with secretory granules during subcellular fractionation of the pancreas and that Sdmg1 and the secretory granule marker Vamp2 are localized to distinct subdomains in the secretory granule membrane. These data suggest that Sdmg1 is a component of regulated secretory granules in exocrine secretory cells and that the developmental regulation of Sdmg1 expression is related to a role for Sdmg1 in post-Golgi membrane trafficking.

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Comparative proteomics of clathrin-coated vesicles

Cited by 151 publications

References 26 publications

Serotonin Transamidates Rab4 and Facilitates Its Binding to the C Terminus of Serotonin Transporter

Serotonin Transamidates Rab4 and Facilitates Its Binding to the C Terminus of Serotonin Transporter

Molecular Structure, Function, and Dynamics of Clathrin-Mediated Membrane Traffic

Sdmg1 is a component of secretory granules in mouse secretory exocrine tissues

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