The outermost N-terminal region of tapasin facilitates folding of major histocompatibility complex class I
Roder, Gustav ; Geironson Ulfsson, Linda LU ; Darabi, Anna LU ; Harndahl, Mikkel ; Schafer-Nielsen, Claus ; Skjodt, Karsten ; Buus, Soren and Paulsson, Kajsa M LU
(2009)
In European Journal of Immunology
39(10).
p.2682-2694
- Abstract
- Tapasin (Tpn) is an ER chaperone that is uniquely dedicated to MHC-I biosynthesis. It binds MHC-I molecules, integrates them into peptide-loading complexes, and exerts quality control of the bound peptides; only when an "optimal peptide" is bound will the MHC-I be released and exported to the cell surface for presentation to T cells. The exact mechanisms of Tpn quality control and the criteria for being an optimal peptide are still unknown. Here, we have generated a recombinant fragment of human Tpn, Tpn(1-87) (representing the 87 N-terminal and ER-luminal amino acids of the mature Tpn protein). Using a biochemical peptide-MHC-I-binding assay, recombinant Tpn(1-87) was found to specifically facilitate peptide-dependent folding of... (More)
- Tapasin (Tpn) is an ER chaperone that is uniquely dedicated to MHC-I biosynthesis. It binds MHC-I molecules, integrates them into peptide-loading complexes, and exerts quality control of the bound peptides; only when an "optimal peptide" is bound will the MHC-I be released and exported to the cell surface for presentation to T cells. The exact mechanisms of Tpn quality control and the criteria for being an optimal peptide are still unknown. Here, we have generated a recombinant fragment of human Tpn, Tpn(1-87) (representing the 87 N-terminal and ER-luminal amino acids of the mature Tpn protein). Using a biochemical peptide-MHC-I-binding assay, recombinant Tpn(1-87) was found to specifically facilitate peptide-dependent folding of HLA-A*0201. Furthermore, we used Tpn(1-87) to generate a monoclonal antibody, alpha Tpn(1-87/80), specific for natural human Tpn and capable of cellular staining of ER localized Tpn. Using overlapping peptides, the epitope of alpha Tpn(1-87)/80 was located to Tpn(40-44), which maps to a surface-exposed loop on the Tpn structure. Together, these results demonstrate that the N-terminal region of Tpn can be recombinantly expressed and adopt a structure, which at least partially resembles that of WT Tpn, and that this region of Tpn features chaperone activity facilitating peptide binding of MHC-I. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1505827
- author
- Roder, Gustav
; Geironson Ulfsson, Linda
LU
; Darabi, Anna
LU
; Harndahl, Mikkel
; Schafer-Nielsen, Claus
; Skjodt, Karsten
; Buus, Soren
and Paulsson, Kajsa M
LU
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Antibodies, Antigen processing, MHC-I
- in
- European Journal of Immunology
- volume
- 39
- issue
- 10
- pages
- 2682 - 2694
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- wos:000271151000007
- scopus:70350547789
- ISSN
- 1521-4141
- DOI
- 10.1002/eji.200939364
- language
- English
- LU publication?
- yes
- id
- e5363cdd-760a-4d92-bd19-e90ecd7a63fd (old id 1505827)
- date added to LUP
- 2016-04-01 11:53:04
- date last changed
- 2022-01-26 19:42:41
@article{e5363cdd-760a-4d92-bd19-e90ecd7a63fd,
abstract = {{Tapasin (Tpn) is an ER chaperone that is uniquely dedicated to MHC-I biosynthesis. It binds MHC-I molecules, integrates them into peptide-loading complexes, and exerts quality control of the bound peptides; only when an "optimal peptide" is bound will the MHC-I be released and exported to the cell surface for presentation to T cells. The exact mechanisms of Tpn quality control and the criteria for being an optimal peptide are still unknown. Here, we have generated a recombinant fragment of human Tpn, Tpn(1-87) (representing the 87 N-terminal and ER-luminal amino acids of the mature Tpn protein). Using a biochemical peptide-MHC-I-binding assay, recombinant Tpn(1-87) was found to specifically facilitate peptide-dependent folding of HLA-A*0201. Furthermore, we used Tpn(1-87) to generate a monoclonal antibody, alpha Tpn(1-87/80), specific for natural human Tpn and capable of cellular staining of ER localized Tpn. Using overlapping peptides, the epitope of alpha Tpn(1-87)/80 was located to Tpn(40-44), which maps to a surface-exposed loop on the Tpn structure. Together, these results demonstrate that the N-terminal region of Tpn can be recombinantly expressed and adopt a structure, which at least partially resembles that of WT Tpn, and that this region of Tpn features chaperone activity facilitating peptide binding of MHC-I.}},
author = {{Roder, Gustav and Geironson Ulfsson, Linda and Darabi, Anna and Harndahl, Mikkel and Schafer-Nielsen, Claus and Skjodt, Karsten and Buus, Soren and Paulsson, Kajsa M}},
issn = {{1521-4141}},
keywords = {{Antibodies; Antigen processing; MHC-I}},
language = {{eng}},
number = {{10}},
pages = {{2682--2694}},
publisher = {{John Wiley & Sons Inc.}},
series = {{European Journal of Immunology}},
title = {{The outermost N-terminal region of tapasin facilitates folding of major histocompatibility complex class I}},
url = {{http://dx.doi.org/10.1002/eji.200939364}},
doi = {{10.1002/eji.200939364}},
volume = {{39}},
year = {{2009}},
}