Secretory proteins in a ciliate cell: identification of epitopes common to numerous polypeptides

Details

Serval ID
serval:BIB_52C8759F5DE9
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Secretory proteins in a ciliate cell: identification of epitopes common to numerous polypeptides
Journal
Biologie Cellulaire
Author(s)
Eperon  S., Peck  R. K., Swiderski  B.
ISSN
0248-4900
Publication state
Published
Issued date
1993
Peer-reviewed
Oui
Volume
78
Number
3
Pages
207-16
Notes
Journal Article
Research Support, Non-U.S. Gov't
Abstract
Secretory vesicles of the ciliate Pseudomicrothorax dubius, called trichocysts, are separated into > 40 proteins by two-dimensional gel electrophoresis. The trichocyst, composed of a shaft and four arms, is in a condensed state when docked in the cell cortex, and it elongates into an extended state during exocytosis. Monoclonal antibodies (mAbs) were raised against trichocyst proteins. Their reactivities were analysed: 1) on Western blots of extended, isolated trichocysts by immunolabeling; 2) on entire cells and extended trichocysts by indirect immunofluorescent binding assay (IFA); 3) on semi-thin sectioned cells by IFA; and 4) on ultra-thin sections of cells by immunogold labeling. mAb IV 4E5 labels major trichocyst proteins at 15-19, 22 and 24 kDa, pI 4.6-6.6. The epitope recognized by mAb IV 4E5 is common to as many as 30 proteins and suggests a family of proteins with possible sequence homology. By IFA, the shafts of extended trichocysts are labeled. The shafts of condensed trichocysts are labeled on both semi-thin sections in Lowicryl and ultrathin sections. On semi-thin Epon sections, the part of the trichocyst which is labeled is arm-like. mAb VI 2D12 labels three major trichocyst proteins at 31 kDa, pI 5.0-5.4. The arms of extended trichocysts are labeled by IFA, but are only weakly labeled on ultrathin sections. The shaft of extended trichocysts is labeled by IFA, and the shaft of condensed trichocysts is labeled on ultrathin sections.
Keywords
Animals Antibodies, Monoclonal Blotting, Western Ciliophora/*ultrastructure Cytoplasmic Granules/*ultrastructure Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Epitopes/*analysis Immunohistochemistry Microscopy, Immunoelectron Molecular Weight Organelles/*ultrastructure Protozoan Proteins/*analysis/isolation & purification
Pubmed
Web of science
Create date
17/01/2008 14:27
Last modification date
20/08/2019 15:08
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