TY - JOUR
T1 - Mechanical properties of BiP protein determined by nano-rheology
AU - Casanova-Morales, Nathalie
AU - Quiroga-Roger, Diego
AU - Alfaro-Valdés, Hilda M.
AU - Alavi, Zahra
AU - Lagos-Espinoza, Miguel I.A.
AU - Zocchi, Giovanni
AU - Wilson, Christian A.M.
N1 - Funding Information:
This study was supported by the Fondo Nacional de Desarrollo Científico y Tecnológico (Fondecyt), Chile, Project 11130263 and 1181361 (CW), Project 3160645 (DQ), Project CONICYT + NERC + Programa de Colaboración Internacional PCI-PII20150073 (CW), U-inicia from the Vicerrectoría de Investigación Universidad de Chile (CW), and NSF grant DMR-1404400 (GZ). We thanks David Shaw King from Howard Hughes Medical Institute and University of California at Berkeley for peptide synthesis; Dr. Mauricio Baez, Maira Rivera and Andrés Bustamante for help in circular dichroism measurements (funded by grant FONDEQUIP EQM140151).
Funding Information:
This study was supported by the Fondo Nacional de Desarrollo Científico y Tecnológico (Fondecyt), Chile, Project 11130263 and 1181361 (CW), Project 3160645 (DQ), Project CONICYT 1 NERC 1 Programa de Colaboración Internacional PCI-PII20150073 (CW), U-inicia from the Vicerrectoría de Investigación Uni-versidad de Chile (CW), and NSF grant DMR-1404400 (GZ). We thanks David Shaw King from Howard Hughes Medical Institute and University of California at Berkeley for peptide synthesis; Dr. Mauricio Baez, Maira Rivera and Andrés Bustamante for help in circular dichroism measurements (funded by grant FONDEQUIP EQM140151).
Funding Information:
Grant sponsor: Vicerrectoría de Investigación, Universidad de Chile; Grant number: U-inicia; Grant sponsor: Fondo de Equipamiento Científico y Tecnológico; Grant number: EQM140151; Grant sponsor: Comisión Nacional de Investigación Científica y Tecnológica; Grant number: Project CONICYT + NERC + Programa de Colaboración; Grant sponsor: National Science Foundation; Grant number: DMR-1404400 (GZ); Grant sponsor: Fondo Nacional de Ciencia Tecnología e Innovación; Grant number: Project 11130263 and 1181361 (CW), Project 3160645 (DQ).
Publisher Copyright:
© 2018 The Protein Society
PY - 2018/8
Y1 - 2018/8
N2 - Immunoglobulin Binding Protein (BiP) is a chaperone and molecular motor belonging to the Hsp70 family, involved in the regulation of important biological processes such as synthesis, folding and translocation of proteins in the Endoplasmic Reticulum. BiP has two highly conserved domains: the N-terminal Nucleotide-Binding Domain (NBD), and the C-terminal Substrate-Binding Domain (SBD), connected by a hydrophobic linker. ATP binds and it is hydrolyzed to ADP in the NBD, and BiP's extended polypeptide substrates bind in the SBD. Like many molecular motors, BiP function depends on both structural and catalytic properties that may contribute to its performance. One novel approach to study the mechanical properties of BiP considers exploring the changes in the viscoelastic behavior upon ligand binding, using a technique called nano-rheology. This technique is essentially a traditional rheology experiment, in which an oscillatory force is directly applied to the protein under study, and the resulting average deformation is measured. Our results show that the folded state of the protein behaves like a viscoelastic material, getting softer when it binds nucleotides- ATP, ADP, and AMP-PNP-, but stiffer when binding HTFPAVL peptide substrate. Also, we observed that peptide binding dramatically increases the affinity for ADP, decreasing it dissociation constant (K D ) around 1000 times, demonstrating allosteric coupling between SBD and NBD domains.
AB - Immunoglobulin Binding Protein (BiP) is a chaperone and molecular motor belonging to the Hsp70 family, involved in the regulation of important biological processes such as synthesis, folding and translocation of proteins in the Endoplasmic Reticulum. BiP has two highly conserved domains: the N-terminal Nucleotide-Binding Domain (NBD), and the C-terminal Substrate-Binding Domain (SBD), connected by a hydrophobic linker. ATP binds and it is hydrolyzed to ADP in the NBD, and BiP's extended polypeptide substrates bind in the SBD. Like many molecular motors, BiP function depends on both structural and catalytic properties that may contribute to its performance. One novel approach to study the mechanical properties of BiP considers exploring the changes in the viscoelastic behavior upon ligand binding, using a technique called nano-rheology. This technique is essentially a traditional rheology experiment, in which an oscillatory force is directly applied to the protein under study, and the resulting average deformation is measured. Our results show that the folded state of the protein behaves like a viscoelastic material, getting softer when it binds nucleotides- ATP, ADP, and AMP-PNP-, but stiffer when binding HTFPAVL peptide substrate. Also, we observed that peptide binding dramatically increases the affinity for ADP, decreasing it dissociation constant (K D ) around 1000 times, demonstrating allosteric coupling between SBD and NBD domains.
KW - BiP chaperone
KW - allosteric communication
KW - binding parameters
KW - mechanical properties
KW - nano-rheology
KW - viscoelasticity
UR - http://www.scopus.com/inward/record.url?scp=85053793667&partnerID=8YFLogxK
U2 - 10.1002/pro.3432
DO - 10.1002/pro.3432
M3 - Article
C2 - 29696702
AN - SCOPUS:85053793667
SN - 0961-8368
VL - 27
SP - 1418
EP - 1426
JO - Protein Science
JF - Protein Science
IS - 8
ER -