Mapping the Ca2 + induced structural change in calreticulin
Boelt, Sanne Grundvad ; Norn, Christoffer LU ; Rasmussen, Morten Ib ; André, Ingemar LU
; Čiplys, Evaldas
; Slibinskas, Rimantas
; Houen, Gunnar
and Højrup, Peter
(2016)
In Journal of Proteomics
142.
p.138-148
- Abstract
Calreticulin is a highly conserved multifunctional protein implicated in many different biological systems and has therefore been the subject of intensive research. It is primarily present in the endoplasmatic reticulum where its main functions are to regulate Ca2 + homeostasis, act as a chaperone and stabilize the MHC class I peptide-loading complex. Although several high-resolution structures of calreticulin exist, these only cover three-quarters of the entire protein leaving the extended structures unsolved. Additionally, the structure of calreticulin is influenced by the presence of Ca2 +. The conformational changes induced by Ca2 + have not been determined yet as they are hard to study with... (More)
Calreticulin is a highly conserved multifunctional protein implicated in many different biological systems and has therefore been the subject of intensive research. It is primarily present in the endoplasmatic reticulum where its main functions are to regulate Ca2 + homeostasis, act as a chaperone and stabilize the MHC class I peptide-loading complex. Although several high-resolution structures of calreticulin exist, these only cover three-quarters of the entire protein leaving the extended structures unsolved. Additionally, the structure of calreticulin is influenced by the presence of Ca2 +. The conformational changes induced by Ca2 + have not been determined yet as they are hard to study with traditional approaches. Here, we investigated the Ca2 +-induced conformational changes with a combination of chemical cross-linking, mass spectrometry, bioinformatics analysis and modelling in Rosetta. Using a bifunctional linker, we found a large Ca2 +-induced change to the cross-linking pattern in calreticulin. Our results are consistent with a high flexibility in the P-loop, a stabilization of the acidic C-terminal and a relatively close interaction of the P-loop and the acidic C-terminal. Biological significance The function of calreticulin, an endoplasmatic reticulin chaperone, is affected by fluctuations in Ca2 + concentration, but the structural mechanism is unknown. The present work suggests that Ca2 +-dependent regulation is caused by different conformations of a long proline-rich loop that changes the accessibility to the peptide/lectin-binding site. Our results indicate that the binding of Ca2 + to calreticulin may thus not only just be a question of Ca2 + storage but is likely to have an impact on the chaperone activity.
(Less)
- author
- Boelt, Sanne Grundvad
; Norn, Christoffer
LU
; Rasmussen, Morten Ib
; André, Ingemar
LU
; Čiplys, Evaldas
; Slibinskas, Rimantas
; Houen, Gunnar
and Højrup, Peter
- organization
- publishing date
- 2016-01-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- BS d, Calreticulin, Chemical cross-linking, Mass spectrometry, MassAI software, Protein structure, Rosetta modelling
- in
- Journal of Proteomics
- volume
- 142
- pages
- 11 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:84973889238
- pmid:27195812
- wos:000378468900015
- ISSN
- 1874-3919
- DOI
- 10.1016/j.jprot.2016.05.015
- language
- English
- LU publication?
- yes
- id
- c33c8e43-a5ea-4c46-881b-15241d4ebd61
- date added to LUP
- 2017-02-15 09:16:37
- date last changed
- 2024-04-14 04:26:41
@article{c33c8e43-a5ea-4c46-881b-15241d4ebd61,
abstract = {{<p>Calreticulin is a highly conserved multifunctional protein implicated in many different biological systems and has therefore been the subject of intensive research. It is primarily present in the endoplasmatic reticulum where its main functions are to regulate Ca<sup>2 +</sup> homeostasis, act as a chaperone and stabilize the MHC class I peptide-loading complex. Although several high-resolution structures of calreticulin exist, these only cover three-quarters of the entire protein leaving the extended structures unsolved. Additionally, the structure of calreticulin is influenced by the presence of Ca<sup>2 +</sup>. The conformational changes induced by Ca<sup>2 +</sup> have not been determined yet as they are hard to study with traditional approaches. Here, we investigated the Ca<sup>2 +</sup>-induced conformational changes with a combination of chemical cross-linking, mass spectrometry, bioinformatics analysis and modelling in Rosetta. Using a bifunctional linker, we found a large Ca<sup>2 +</sup>-induced change to the cross-linking pattern in calreticulin. Our results are consistent with a high flexibility in the P-loop, a stabilization of the acidic C-terminal and a relatively close interaction of the P-loop and the acidic C-terminal. Biological significance The function of calreticulin, an endoplasmatic reticulin chaperone, is affected by fluctuations in Ca<sup>2 +</sup> concentration, but the structural mechanism is unknown. The present work suggests that Ca<sup>2 +</sup>-dependent regulation is caused by different conformations of a long proline-rich loop that changes the accessibility to the peptide/lectin-binding site. Our results indicate that the binding of Ca<sup>2 +</sup> to calreticulin may thus not only just be a question of Ca<sup>2 +</sup> storage but is likely to have an impact on the chaperone activity.</p>}},
author = {{Boelt, Sanne Grundvad and Norn, Christoffer and Rasmussen, Morten Ib and André, Ingemar and Čiplys, Evaldas and Slibinskas, Rimantas and Houen, Gunnar and Højrup, Peter}},
issn = {{1874-3919}},
keywords = {{BS d; Calreticulin; Chemical cross-linking; Mass spectrometry; MassAI software; Protein structure; Rosetta modelling}},
language = {{eng}},
month = {{01}},
pages = {{138--148}},
publisher = {{Elsevier}},
series = {{Journal of Proteomics}},
title = {{Mapping the Ca<sup>2 +</sup> induced structural change in calreticulin}},
url = {{http://dx.doi.org/10.1016/j.jprot.2016.05.015}},
doi = {{10.1016/j.jprot.2016.05.015}},
volume = {{142}},
year = {{2016}},
}