Abstract:
:The surface coat of procyclic forms of Trypanosoma brucei consists of related, internally repetitive glycoproteins known as EP and GPEET procyclins. Previously we showed that the extracellular domain of GPEET is phosphorylated. We now show that phosphorylation of this glycosylphosphatidylinositol-anchored surface protein can be induced in vitro using a procyclic membrane extract. Using antibodies that recognize either the phosphorylated or unphosphorylated form of GPEET, we analyzed their expression during differentiation of bloodstream forms to procyclic forms. Unphosphorylated GPEET, together with EP, was detected in cell lysates 2-4 hours after initiating differentiation whereas phosphorylated GPEET only appeared after 24 hours. Surface expression of EP and both forms of GPEET occurred after 24-48 hours and correlated with the detection of phosphorylated GPEET on immuno-blots. Electron micrographs showed that unphosphorylated GPEET was predominantly in the flagellar pocket whereas the phosphorylated form was distributed over the cell surface. In contrast, expression of a membrane-bound human placental alkaline phosphatase in procyclic forms caused the accumulation of dephosphorylated GPEET on the cell surface, while the phosphorylated form was restricted to the flagellar pocket. A GPEET-Fc fusion protein, which was retained intracellularly, was not phosphorylated. We propose that unphosphorylated GPEET procyclin is transported to a location close to or at the cell surface, most probably the flagellar pocket, where it becomes phosphorylated. To the best of our knowledge, this study represents the first localization of phosphorylated and unphosphorylated forms of a GPI-anchored protein within a cell.
journal_name
J Cell Scijournal_title
Journal of cell scienceauthors
Bütikofer P,Vassella E,Ruepp S,Boschung M,Civenni G,Seebeck T,Hemphill A,Mookherjee N,Pearson TW,Roditi Ikeywords:
subject
Has Abstractpub_date
1999-06-01 00:00:00pages
1785-95eissn
0021-9533issn
1477-9137journal_volume
112 ( Pt 11)pub_type
杂志文章abstract::The adaptive immune response depends on the interaction of T cells and antigen-presenting cells at the immune synapse. Formation of the immune synapse and the subsequent T-cell activation are highly dependent on the actin cytoskeleton. In this work, we describe that T cells express drebrin, a neuronal actin-binding pr...
journal_title:Journal of cell science
pub_type: 杂志文章
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abstract::A heterogenous population of intact chondrons extracted from low-speed homogenates of canine tibial cartilage were stained by indirect immunofluorescence methods with a polyclonal antibody to type VI collagen. In each of the four chondron groups examined, anti-(type VI collagen) anti-serum was concentrated in the caps...
journal_title:Journal of cell science
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journal_title:Journal of cell science
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journal_title:Journal of cell science
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journal_title:Journal of cell science
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journal_title:Journal of cell science
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pub_type: 杂志文章,评审
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journal_title:Journal of cell science
pub_type: 杂志文章
doi:
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journal_title:Journal of cell science
pub_type: 杂志文章
doi:
更新日期:1993-03-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章
doi:
更新日期:1989-10-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章
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更新日期:1992-03-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
pub_type: 杂志文章
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更新日期:2000-05-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章,评审
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更新日期:2004-02-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章
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更新日期:2000-03-01 00:00:00
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
pub_type: 杂志文章
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更新日期:1993-05-01 00:00:00
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journal_title:Journal of cell science
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journal_title:Journal of cell science
pub_type: 杂志文章
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journal_title:Journal of cell science
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journal_title:Journal of cell science
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