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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Conformational flexibility of GRASPs and their constituent PDZ subdomains reveals structural basis of their promiscuous interactome

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Author(s):
Mendes, Luis Felipe S. [1, 2] ; Batista, Mariana R. B. [2] ; Judge, Peter J. [1] ; Watts, Anthony [1] ; Redfield, Christina [1] ; Costa-Filho, Antonio J. [2]
Total Authors: 6
Affiliation:
[1] Univ Oxford, Dept Biochem, Oxford - England
[2] Univ Sao Paulo, Mol Biophys Lab, Ribeirao Preto Sch Philosophy Sci & Literature, Phys Dept, Ribeirao Preto - Brazil
Total Affiliations: 2
Document type: Journal article
Source: FEBS Journal; v. 287, n. 15 JAN 2020.
Web of Science Citations: 0
Abstract

The Golgi complex is a central component of the secretory pathway, responsible for several critical cellular functions in eukaryotes. The complex is organized by the Golgi matrix that includes the Golgi reassembly and stacking protein (GRASP), which was shown to be involved in cisternae stacking and lateral linkage in metazoan. GRASPs also have critical roles in other processes, with an unusual ability to interact with several different binding partners. The conserved N terminus of the GRASP family includes two PSD-95, DLG, and ZO-1 (PDZ) domains. Previous crystallographic studies of orthologues suggest that PDZ1 and PDZ2 have similar conformations and secondary structure content. However, PDZ1 alone mediates nearly all interactions between GRASPs and their partners. In this work, NMR, synchrotron radiation CD, and molecular dynamics (MD) were used to examine the structure, flexibility, and stability of the two constituent PDZ domains. GRASP PDZs are structured in an unusual beta(3)alpha(1)beta(4)beta(5)alpha(2)beta(6)beta(1)beta(2) secondary structural arrangement and NMR data indicate that the PDZ1 binding pocket is formed by a stable beta(2)-strand and a more flexible and unstable alpha(2)-helix, suggesting an explanation for the higher PDZ1 promiscuity. The conformational free energy profiles of the two PDZ domains were calculated using MD simulations. The data suggest that, after binding, the protein partner significantly reduces the conformational space that GRASPs can access by stabilizing one particular conformation, in a partner-dependent fashion. The structural flexibility of PDZ1, modulated by PDZ2, and the coupled, coordinated movement between the two PDZs enable GRASPs to interact with multiple partners, allowing them to function as promiscuous, multitasking proteins. (AU)

FAPESP's process: 12/20367-3 - Structural and functional studies of the Golgi Re-Assembly and Stacking Protein (GRASP) from Cryptococcus neoformans
Grantee:Antonio José da Costa Filho
Support Opportunities: Regular Research Grants
FAPESP's process: 15/50366-7 - Resolving mechanistic details of peptide transport across membranes using crystallographic and non-crystallographic structural biology approaches
Grantee:Antonio José da Costa Filho
Support Opportunities: Regular Research Grants
FAPESP's process: 16/09676-5 - Nuclear magnetic resonance studies of the Golgi reassembly and stacking protein from Cryptococcus neoformans
Grantee:Luis Felipe Santos Mendes
Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
FAPESP's process: 17/24669-8 - Unraveling the molecular bases of the early protein secretory pathway in humans using biophysical techniques
Grantee:Luis Felipe Santos Mendes
Support Opportunities: Scholarships in Brazil - Post-Doctoral