scholarly journals Sequences within the Cytoplasmic Domain of Gp180/Carboxypeptidase D Mediate Localization to the Trans-Golgi Network

1999 ◽  
Vol 10 (1) ◽  
pp. 35-46 ◽  
Author(s):  
Francis J. Eng ◽  
Oleg Varlamov ◽  
Lloyd D. Fricker
Keyword(s):  
Cell Surface ◽  
Secretory Pathway ◽  
Point Mutations ◽  
Cell Surface Receptor ◽  
Type I ◽  
Multiple Sequence ◽  
Trans Golgi Network ◽  
Duck Hepatitis ◽  
Surface Receptor ◽  
Golgi Network

Gp180, a duck protein that was proposed to be a cell surface receptor for duck hepatitis B virus, is the homolog of metallocarboxypeptidase D, a mammalian protein thought to function in the trans-Golgi network (TGN) in the processing of proteins that transit the secretory pathway. Both gp180 and mammalian metallocarboxypeptidase D are type I integral membrane proteins that contain a 58-residue cytosolic C-terminal tail that is highly conserved between duck and rat. To investigate the regions of the gp180 tail involved with TGN retention and intracellular trafficking, gp180 and various deletion and point mutations were expressed in the AtT-20 mouse pituitary corticotroph cell line. Full length gp180 is enriched in the TGN and also cycles to the cell surface. Truncation of the C-terminal 56 residues of the cytosolic tail eliminates the enrichment in the TGN and the retrieval from the cell surface. Truncation of 12–43 residues of the tail reduced retention in the TGN and greatly accelerated the turnover of the protein. In contrast, deletion of the C-terminal 45 residues, which truncates a potential YxxL-like sequence (FxxL), reduced the protein turnover and caused accumulation of the protein on the cell surface. A point mutation of the FxxL sequence to AxxL slowed internalization, showing that this element is important for retrieval from the cell surface. Mutation of a pair of casein kinase II sites within an acidic cluster showed that they are also important for trafficking. The present study demonstrates that multiple sequence elements within the cytoplasmic tail of gp180 participate in TGN localization.

The steady state distribution of humTGN46 is not significantly altered in cells defective in clathrin-mediated endocytosis

1998 ◽  
Vol 111 (23) ◽  
pp. 3451-3458 ◽  
Author(s):  
G. Banting ◽  
R. Maile ◽  
E.P. Roquemore
Keyword(s):  
Steady State ◽  
Cell Surface ◽  
Dominant Negative ◽  
Type I ◽  
Steady State Distribution ◽  
State Distribution ◽  
Trans Golgi Network ◽  
Defective Mutant ◽  
Dynamin I ◽  
Golgi Network

It has been shown previously that whilst the rat type I integral membrane protein TGN38 (ratTGN38) is predominantly localised to the trans-Golgi network this protein does reach the cell surface from where it is internalised and delivered back to the trans-Golgi network. This protein thus provides a suitable tool for the investigation of trafficking pathways between the trans-Golgi network and the cell surface and back again. The human orthologue of ratTGN38, humTGN46, behaves in a similar fashion. These proteins are internalised from the cell surface via clathrin mediated endocytosis, a process which is dependent upon the GTPase activity of dynamin. We thus reasoned that humTGN46 would accumulate at the surface of cells rendered defective in clathrin mediated endocytosis by virtue of the fact that they express a GTPase defective mutant of dynamin I. It did not. In fact, expression of a dominant negative GTPase defective mutant of dynamin I had no detectable effect on the steady state distribution of humTGN46. One explanation for this observation is that humTGN46 does not travel directly to the cell surface from the trans-Golgi network. Further studies on cells expressing the dominant negative GTPase defective mutant of dynamin I indicate that the major recycling pathway for humTGN46 is in fact between the trans-Golgi network and the early endosome.

Dynamic palmitoylation of lymphoma proprotein convertase prolongs its half-life, but is not essential for trans-Golgi network localization

2000 ◽  
Vol 352 (3) ◽  
pp. 827-833 ◽  
Author(s):  
Jan-Willem H. P. VAN DE LOO ◽  
Meike TEUCHERT ◽  
Ilse PAULI ◽  
Evelyn PLETS ◽  
Wim J. M.VAN DE VEN ◽  
...  
Keyword(s):  
Half Life ◽  
Secretory Pathway ◽  
Transmembrane Protein ◽  
Brefeldin A ◽  
Cytoplasmic Tail ◽  
Cysteine Residue ◽  
Type I ◽  
Proprotein Convertase ◽  
Trans Golgi Network ◽  
Golgi Network

Proprotein convertases are responsible for the endoproteolytic activation of proproteins in the secretory pathway. The most recently discovered member of this family, lymphoma proprotein convertase (LPC), is a type-I transmembrane protein. Previously, we have demonstrated that its cytoplasmic tail is palmitoylated. In this study, we have identified the two most proximal cysteine residues in the cytoplasmic tail as palmitoylation sites. Substitution of either cysteine residue by alanine interfered with palmitoylation of the other. Palmitoylation of LPC was found to be sensitive to the protein palmitoyltransferase inhibitor tunicamycin but not cerulenin. It was also insensitive to the drugs brefeldin A, monensin and cycloheximide, indicating that the modification occurs in a late exocytic or endocytic compartment. Turnover of palmitoylated LPC is significantly faster (t1/2 ≈ 50min) than that of the LPC polypeptide backbone (t1/2 ≈ 3h), suggesting that palmitoylation is reversible. Abrogation of palmitoylation reduced the half-life of the LPC protein, but did not affect steady-state localization of LPC in the trans-Golgi network. Finally, LPC could not be detected in detergent-resistant membrane rafts. Taken together, these results suggest that dynamic palmitoylation of LPC is important for stability, but does not function as a dominant trafficking signal.

Intracellular trafficking of metallocarboxypeptidase D in AtT-20 cells: localization to the trans-Golgi network and recycling from the cell surface

1998 ◽  
Vol 111 (7) ◽  
pp. 877-885 ◽  
Author(s):  
O. Varlamov ◽  
L.D. Fricker
Keyword(s):  
Cell Surface ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Secretory Pathway ◽  
Brefeldin A ◽  
Full Length ◽  
Recycling Endosomes ◽  
Trans Golgi Network ◽  
Cell Processes ◽  
Golgi Network

Carboxypeptidase D (CPD) is a recently discovered membrane-bound metallocarboxypeptidase that has been proposed to be involved in the post-translational processing of peptides and proteins that transit the secretory pathway. In the present study, the intracellular distribution of CPD was examined in AtT-20 cells, a mouse anterior pituitary-derived corticotroph. Antisera to CPD stain the same intracellular structures as those labeled with furin and wheat germ agglutinin. This distribution is distinct from carboxypeptidase E, which is localized to the secretory vesicles in the cell processes. The perinuclear distribution of CPD is detected even when the AtT-20 cells are treated with brefeldin A for 1–30 minutes, suggesting that CPD is present in the trans-Golgi network (TGN). Although CPD is predominantly found in the TGN, an antiserum to the full length protein is internalized within 15–30 minutes of incubation at 37 degrees C. In contrast, an antiserum raised against the C-terminal region of CPD does not become internalized, suggesting that this domain is cytosolic. The antiserum to the full length CPD is internalized to a structure that co-stains with furin and wheat germ agglutinin, but is distinct from transferrin recycling endosomes. The internalization of CPD is not substantially affected by treatment of the AtT-20 cells with brefeldin A. These data are consistent with the cycling of CPD to the cell surface and back to the TGN. The TGN localization of CPD raises the possibility of a role for this enzyme in the processing of proteins that transit the secretory pathway.

scholarly journals The Arabidopsis receptor kinase STRUBBELIG undergoes clathrin-dependent endocytosis

2019 ◽  
Vol 70 (15) ◽  
pp. 3881-3894 ◽  
Author(s):  
Jin Gao ◽  
Ajeet Chaudhary ◽  
Prasad Vaddepalli ◽  
Marie-Kristin Nagel ◽  
Erika Isono ◽  
...  
Keyword(s):  
Cell Surface ◽  
Fluorescent Protein ◽  
Growth Patterns ◽  
Cell Surface Receptor ◽  
Receptor Kinase ◽  
Surface Receptors ◽  
Surface Receptor ◽  
Green Fluorescent ◽  
Golgi Network

AbstractSignaling mediated by cell surface receptor kinases is central to the coordination of growth patterns during organogenesis. Receptor kinase signaling is in part controlled through endocytosis and subcellular distribution of the respective receptor kinase. For the majority of plant cell surface receptors, the underlying trafficking mechanisms are not characterized. In Arabidopsis, tissue morphogenesis requires the atypical receptor kinase STRUBBELIG (SUB). Here, we studied the endocytic mechanism of SUB. Our data revealed that a functional SUB–enhanced green fluorescent protein (EGFP) fusion is ubiquitinated in vivo. We further showed that plasma membrane-bound SUB:EGFP becomes internalized in a clathrin-dependent fashion. We also found that SUB:EGFP associates with the trans-Golgi network and accumulates in multivesicular bodies and the vacuole. Co-immunoprecipitation experiments revealed that SUB:EGFP and clathrin are present within the same protein complex. Our genetic analysis showed that SUB and CLATHRIN HEAVY CHAIN (CHC) 2 regulate root hair patterning. By contrast, genetic reduction of CHC activity ameliorates the floral defects of sub mutants. Taken together, the data indicate that SUB undergoes clathrin-mediated endocytosis, that this process does not rely on stimulation of SUB signaling by an exogenous agent, and that SUB genetically interacts with clathrin-dependent pathways in a tissue-specific manner.

scholarly journals A cell surface receptor complex for collagen type I recognizes the Arg-Gly-Asp sequence.

1987 ◽  
Vol 104 (3) ◽  
pp. 585-593 ◽  
Author(s):  
S Dedhar ◽  
E Ruoslahti ◽  
M D Pierschbacher
Keyword(s):  
Cell Surface ◽  
Type I Collagen ◽  
Attachment Site ◽  
Cell Surface Receptor ◽  
Type I ◽  
Receptor Complex ◽  
Collagen Binding ◽  
Human Osteosarcoma Cells ◽  
Surface Receptor ◽  
A Cell

To isolate collagen-binding cell surface proteins, detergent extracts of surface-iodinated MG-63 human osteosarcoma cells were chromatographed on affinity matrices of either type I collagen-Sepharose or Sepharose carrying a collagen-like triple-helical peptide. The peptide was designed to be triple helical and to contain the sequence Arg-Gly-Asp, which has been implicated as the cell attachment site of fibronectin, vitronectin, fibrinogen, and von Willebrand factor, and is also present in type I collagen. Three radioactive polypeptides having apparent molecular masses of 250 kD, 70 kD, and 30 kD were distinguishable in that they showed affinity toward the collagen and collagen-like peptide affinity columns, and could be specifically eluted from these columns with a solution of an Arg-Gly-Asp-containing peptide, Gly-Arg-Gly-Asp-Thr-Pro. These collagen-binding polypeptides associated with phosphatidylcholine liposomes, and the resulting liposomes bound specifically to type I collagen or the collagen-like peptide but not to fibronectin or vitronectin or heat-denatured collagen. The binding of these liposomes to type I collagen could be inhibited with the peptide Gly-Arg-Gly-Asp-Thr-Pro and with EDTA, but not with a variant peptide Gly-Arg-Gly-Glu-Ser-Pro. We conclude from these data that these three polypeptides are membrane molecules that behave as a cell surface receptor (or receptor complex) for type I collagen by interacting with it through the Arg-Gly-Asp tripeptide adhesion signal. The lack of binding to denatured collagen suggests that the conformation of the Arg-Gly-Asp sequence is important in the recognition of collagen by the receptor complex.

scholarly journals The Combination of Type I IFN, TNF-α, and Cell Surface Receptor Engagement with Dendritic Cells Enables NK Cells To Overcome Immune Evasion by Dengue Virus

2014 ◽  
Vol 193 (10) ◽  
pp. 5065-5075 ◽  
Author(s):  
Daniel Say Liang Lim ◽  
Nobuyo Yawata ◽  
Kevin John Selva ◽  
Na Li ◽  
Chen Yu Tsai ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Cell Surface ◽  
Dengue Virus ◽  
Nk Cells ◽  
Immune Evasion ◽  
Cell Surface Receptor ◽  
Type I ◽  
Type I Ifn ◽  
Tnf Α ◽  
Surface Receptor

scholarly journals Dual pulse-chase microscopy reveals early divergence in the biosynthetic trafficking of the Na,K-ATPase and E-cadherin

2015 ◽  
Vol 26 (24) ◽  
pp. 4401-4411 ◽  
Author(s):  
Glen A. Farr ◽  
Michael Hull ◽  
Emily H. Stoops ◽  
Rosalie Bateson ◽  
Michael J. Caplan
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Golgi Complex ◽  
Plasma Membranes ◽  
Secretory Pathway ◽  
Trans Golgi Network ◽  
Polarized Epithelial Cells ◽  
Golgi Network ◽  
E Cadherin

Recent evidence indicates that newly synthesized membrane proteins that share the same distributions in the plasma membranes of polarized epithelial cells can pursue a variety of distinct trafficking routes as they travel from the Golgi complex to their common destination at the cell surface. In most polarized epithelial cells, both the Na,K-ATPase and E-cadherin are localized to the basolateral domains of the plasma membrane. To examine the itineraries pursued by newly synthesized Na,K-ATPase and E-cadherin in polarized MDCK epithelial cells, we used the SNAP and CLIP labeling systems to fluorescently tag temporally defined cohorts of these proteins and observe their behaviors simultaneously as they traverse the secretory pathway. These experiments reveal that E-cadherin is delivered to the cell surface substantially faster than is the Na,K-ATPase. Furthermore, the surface delivery of newly synthesized E-cadherin to the plasma membrane was not prevented by the 19°C temperature block that inhibits the trafficking of most proteins, including the Na,K-ATPase, out of the trans-Golgi network. Consistent with these distinct behaviors, populations of newly synthesized E-cadherin and Na,K-ATPase become separated from one another within the trans-Golgi network, suggesting that they are sorted into different carrier vesicles that mediate their post-Golgi trafficking.

scholarly journals Identification of integrin alpha 2 beta 1 as cell surface receptor for the carboxyl-terminal propeptide of type I procollagen.

1994 ◽  
Vol 269 (33) ◽  
pp. 20982-20986
Author(s):  
S.A. Weston ◽  
D.J. Hulmes ◽  
A.P. Mould ◽  
R.B. Watson ◽  
M.J. Humphries
Keyword(s):  
Cell Surface ◽  
Cell Surface Receptor ◽  
Type I ◽  
Type I Procollagen ◽  
Surface Receptor ◽  
Carboxyl Terminal

Expression, function, and localization of the REG cell surface receptor

2001 ◽  
Vol 120 (5) ◽  
pp. A18-A19
Author(s):  
B DIECKGRAEFE ◽  
C HOUCHEN ◽  
H ZHANG
Keyword(s):  
Cell Surface ◽  
Cell Surface Receptor ◽  
Surface Receptor
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