Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0038362 (stomatitis)
 8,852 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The fusion of lipid enveloped viruses with cellular membranes is thought to be mediated by the insertion into the target membrane of the N-terminal polypeptides of viral spike glycoproteins. Since membrane destabilization is a necessary step in membrane fusion, we investigated whether synthetic peptides with amino acid sequences corresponding to the N-termini of influenza virus hemagglutinin (HA2), vesicular stomatitis virus G-protein and Sendai virus F-protein, induce the destabilization and fusion of phospholipid vesicles. Membrane destabilization by the peptides was monitored by the release of aqueous contents of large unilamellar phospholipid vesicles. Aggregation was detected by a resonance energy transfer assay. Membrane fusion was followed by means of assays for the intermixing of phospholipids and of aqueous contents. The 17-amino acid HA2 peptide (HA2.17) destabilized phosphatidylcholine (PC) vesicles even at neutral pH, but the rate and extent of destabilization increased at lower pH. This peptide did not mediate appreciable release of contents from phosphatidylserine (PS) vesicles. HA2.17 induced neither aggregation nor fusion of PC or PS vesicles. In contrast, the 7-amino acid N-terminal peptide of G-protein (G.7) destabilized PS-containing membranes and not pure PC vesicles. Although G.7 caused aggregation of and lipid mixing between PS vesicles, it did not mediate any detectable intermixing of aqueous contents. The presence of cholesterol in PC membranes did not affect the destabilization caused by the N-terminal peptide of Sendai virus F-protein (F1.7), suggesting that cholesterol is not necessary for the effective interaction of this peptide with membranes, contrary to earlier proposals. Our results support the hypothesis that the hydrophobic N-terminal region of certain viral envelope proteins insert into and destabilize target membranes.
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PMID:Membrane destabilization by N-terminal peptides of viral envelope proteins. 132 86

Microtubules have been implicated in the transport of vesicles carrying newly synthesized proteins from the trans-Golgi network (TGN) to the cell surface. We have established a quantitative in vitro binding assay to investigate the putative interaction between these exocytic carrier vesicles and the microtubules at the molecular level. TGN-derived exocytic carrier vesicles, labeled with C6NBD-ceramide metabolites or viral glycoproteins, were obtained from polarized filter-grown MDCK II cells by perforation of the apical membrane with a nitrocellulose filter. These exocytic vesicles were incubated with taxol-polymerized tubulin and cytosol, layered on top of a 30% sucrose cushion and subjected to centrifugation. Quantitation of vesicles co-sedimenting with microtubules was done by measuring NBD-fluorescence of viral glycoproteins in the pellet and supernatant fractions. About 25% of the label sedimented through the cushion in the presence of microtubules and cytosol. Both apically and basolaterally targetted carrier vesicles containing influenza virus HA2 or vesicular stomatitis virus G protein, respectively, associated with the microtubules. Only 2-5% NBD-fluorescence was obtained in the pellet when no cytosol or microtubules were added to the vesicles. Negative-stain electron microscopy of resuspended pellets showed distinct microtubule-vesicle complexes. Heat inactivation or treatment of cytosol with N-ethylmaleimide (NEM), or trypsinization of vesicles inhibited the binding of vesicles to microtubules. Furthermore, coating of microtubules with brain microtubule-associated proteins abolished binding. These data suggest that NEM-sensitive cytosolic proteins are required for microtubule-vesicle association, and that the vesicles are bound via trypsin-sensitive receptor proteins on their surface.
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PMID:Binding of exocytic vesicles from MDCK cells to microtubules in vitro. 238 28

Three chimeric proteins were obtained by fusing together the dianthin gene and DNA fragments encoding for the following membrane-acting peptides: the N-terminus of protein G of the vesicular stomatitis virus (KFT25), the N terminus of the HA2 hemagglutinin of influenza virus (pHA2), and a membrane-acting peptide (pJVE). Chimeric dianthins (KFT25DIA, pHA2DIA and pJVEDIA) retained full enzymatic activity in cell-free assays and showed increased ability to induce pH-dependent calcein release from large unilamellar vesicles (LUVs). pHA2DIA and pJVEDIA also showed faster kinetics of interaction with LUVs, while KFT25DIA and pHA2DIA displayed a reduced cytotoxicity as compared to wild-type dianthin. Conjugates made by chemically cross-linking KFT25DIA or pJVEDIA and human transferrin (Tfn) showed greater cell-killing efficiency than conjugates of Tfn and wild-type dianthin. As a consequence, by fusion of membrane-acting peptides to the dianthin sequence the specificity factor (i.e., the ratio between non-specific and specific toxicity) of Tfn-KFT25DIA, Tfn-pHA2DIA and Tfn-pJVEDIA was increased with respect to that of Tfn-based conjugates made with wild-type dianthin. Taken together, our results suggest that genetic fusion of membrane-acting peptides to enzymatic cytotoxins results in the acquisition of new physico-chemical properties exploitable for designing new recombinant cytotoxins and to tackle cell-intoxication mechanisms.
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PMID:Genetic grafting of membrane-acting peptides to the cytotoxin dianthin augments its ability to de-stabilize lipid bilayers and enhances its cytotoxic potential as the component of transferrin-toxin conjugates. 1079 75