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| MATCH THE LOGICAL FALLACY TO THE CORRECT DEFINITION. | 23 hours ago · Leishmania HASPB is a lipoprotein that is exported to the extracellular space from both Leishmania parasites and mammalian cells via an unconventional secretory pathway. Exported. 20 hours ago · Leishmania HASPB is a lipoprotein that is exported to the extracellular space from both Leishmania parasites and mammalian cells via an unconventional secretory pathway. ExportedMissing: mcat. 1 day ago · Leishmania HASPB is a lipoprotein that is exported to the extracellular space from both Leishmania parasites and mammalian cells via an unconventional secretory pathway. Exported. |
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clathrin mcat
Advanced Search Leishmania HASPB is a lipoprotein that is exported to the extracellular space from both Leishmania parasites and mammalian cells via clathirn unconventional secretory clathrin mcat. A detailed characterization of such a CHO mutant cell line revealed that the expression level of the HASPB reporter molecule is unchanged compared to CHO wild-type cells; that it is both myristoylated and palmitoylated; and that it is mainly localized to the plasma membrane as judged by confocal microscopy and subcellular fractionation.
However, based on a quantitative flow cytometry assay clathrin mcat a biochemical biotinylation assay of https://digitales.com.au/blog/wp-content/custom/african-slaves-during-the-nineteenth-century/of-mice-and-men-ban.php proteins, HASPB transport to the outer leaflet of the plasma membrane is largely reduced in this mutant. From these data, we conclude that the subcellular site of HASPB membrane translocation is the plasma membrane as the reporter molecule accumulates in this location when export is blocked.
In a second step, the plasma membrane-resident machinery, which is ,cat disrupted clathrin mcat the CHO mutant cell line, mediates membrane translocation of HASPB. Intriguingly, the angiogenic growth factor FGF-2, another protein secreted by unconventional means, is shown to be secreted normally from the HASPB export mutant cell line.
Materials and Methods
These observations demonstrate that the export machinery component defective in the export mutant cell line functions specifically in the HASPB export pathway. In the case of soluble factors, the principal targeting motifs are N-terminal signal peptides that direct classical secretory proteins to the translocation machinery of the ER Keenan et al. Among these, the most prominent examples are clathrin mcat angiogenic growth factors FGF-1 Jackson et al. Leishmania HASPB is another interesting clathrin mcat of unconventional secretory mcqt that is a component of the surface coat of Leishmania parasites Flinn et al.
Introduction
HASPB is exclusively expressed in infective parasites in both extracellular metacyclics and intracellular amastigotes of L. Therefore, even though palmitoylacyltransferase activities have not only been localized to the Golgi but also to the ER and plasma membranes reviewed by Bijlmakers and Marsh,it is likely that transfer of palmitate to the SH4 domain of HASPB occurs at the Golgi. Following palmitoylation, HASPB is transferred to the plasma membrane but the mechanism of this transport process is unknown.
Based on click the following article considerations, HASPB trafficking appears to be distinct from all three known classes of plasma membrane-targeted proteins carrying dual acylation motifs at their N-termini Bijlmakers and Marsh, The clathrin mcat of these, the Src kinase Lck, is palmitoylated at the Golgi coathrin by brefeldin A-sensitive transport to the plasma membrane Bijlmakers and Marsh, ; Bijlmakers and Marsh, Finally, a third mechanism of plasma membrane targeting of Clathrin mcat acylated proteins is exemplified by the Src kinase Fyn that does not appear to contact intracellular membranes but rather is directly targeted from the cytoplasm to the plasma membrane.
As indicated above, current knowledge clathrin mcat HASPB targeting to the plasma membrane does not fit with any of these examples as HASPB is transiently associated with, and most likely palmitoylated at, the Golgi followed by plasma membrane targeting in a brefeldin A-insensitive manner Denny et al. Based on these mcag, many questions about the molecular mechanism of HASPB targeting to the plasma membrane remain unresolved.
There is also a complete lack of knowledge about the subcellular site of clathrin mcat translocation of HASPB, a process that eventually allows HASPB exposure on the cell surface of eukaryotic cells. In the current study, we introduce a novel experimental system that permits the precise quantification of HASPB export from mammalian cells based on flow cytometry.
Based on clathrin mcat detailed biochemical and morphological characterization of a mutant with this phenotype, we conclude that the site of HASPB membrane translocation is the plasma membrane.
Materials and Methods
These data suggest that in the CHO mutant described, a component of a plasma membrane-resident machinery has been disrupted causing the fully acylated HASPB-GFP reporter molecule to accumulate in the inner leaflet of the plasma membrane. These results support a two-step process clathrin mcat HASPB biogenesis, in which dual acylation of the HASPB SH4 domain is required for delivery to mcxt inner leaflet of the plasma membrane and subsequent recognition by a putative translocation machinery. Intriguingly, the component of click HASPB export apparatus that has been disrupted in the mutant cell line appears to be specific for this export pathway, as the angiogenic growth factor FGF-2, another protein secreted by unconventional means, clathrin mcat found to be exported normally from the HASPB export mutant cell line. Antibodies directed against the transferrin receptor were from Zymed, antibodies directed against GM were purchased from BD Transduction Laboratories.
All constructs were cloned clathrin mcat the retroviral clathrih vector pREV-TRE2 that contains a doxicycline-dependent promoter.]
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