Interaction between a Ca2+-binding protein calreticulin and perforin, a component of the cytotoxic T-cell granules

Details

Serval ID
serval:BIB_3F9660E4B1CE
Type
Article: article from journal or magazin.
Collection
Publications
Title
Interaction between a Ca2+-binding protein calreticulin and perforin, a component of the cytotoxic T-cell granules
Journal
Biochemistry
Author(s)
Andrin  C., Pinkoski  M. J., Burns  K., Atkinson  E. A., Krahenbuhl  O., Hudig  D., Fraser  S. A., Winkler  U., Tschopp  J., Opas  M., Bleackley  R. C., Michalak  M.
ISSN
0006-2960 (Print)
Publication state
Published
Issued date
07/1998
Volume
37
Number
29
Pages
10386-94
Notes
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S. --- Old month value: Jul 21
Abstract
Calreticulin is a component of cytotoxic T-lymphocyte and NK lymphocyte granules. We report here that granule-associated calreticulin terminates with the KDEL endoplasmic reticulum retrieval amino acid sequence and somehow escapes the KDEL retrieval system. In perforin knock-out mice calreticulin is still targeted into the granules. Thus, calreticulin will traffic without perforin to cytotoxic granules. In the granules, calreticulin and perforin are associated as documented by (i) copurification of calreticulin with perforin but not with granzymes and (ii) immunoprecipitation of a calreticulin-perforin complex using specific antibodies. By using calreticulin affinity chromatography and protein ligand blotting we show that perforin binds to calreticulin in the absence of Ca2+ and the two proteins dissociate upon exposure to 0.1 mM or higher Ca2+ concentration. Perforin interacts strongly with the P-domain of calreticulin (the domain which has high Ca2+-binding affinity and chaperone function) as revealed by direct protein-protein interaction, ligand blotting, and the yeast two-hybrid techniques. Our results suggest that calreticulin may act as Ca2+-regulated chaperone for perforin. This action will serve to protect the CTL during biogenesis of granules and may also serve to regulate perforin lytic action after release.
Keywords
Amino Acid Sequence Animals Calcium/physiology Calcium-Binding Proteins/genetics/immunology/*metabolism Calreticulin Cell Line Cytoplasmic Granules/chemistry/immunology/*metabolism Genes, Reporter Membrane Glycoproteins/genetics/immunology/*metabolism Mice Mice, Inbred C57BL Mice, Knockout Molecular Sequence Data Peptide Fragments/metabolism Pore Forming Cytotoxic Proteins Proline/metabolism Protein Structure, Tertiary Rats Recombinant Fusion Proteins/metabolism Ribonucleoproteins/genetics/immunology/*metabolism Saccharomyces cerevisiae/genetics/metabolism T-Lymphocytes, Cytotoxic/*metabolism
Pubmed
Web of science
Create date
24/01/2008 16:18
Last modification date
20/08/2019 14:36
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