Structural and thermodynamic analyses of human TMED1 (p24 gamma 1) Golgi dynamics

Author(s)

Mota, Danielly C. A. M. ; Cardoso, Iara A. ; Mori, Renan M. ; Batista, Mariana R. B. ; Basso, Luis G. M. ; Nonato, M. Cristina ; Costa-Filho, Antonio J. ; Mendes, Luis F. S.

• Document type: Journal article
• Source: Biochimie; v. 192, p. 11-pg., 2022-01-01.

Abstract

The transmembrane emp24 domain-containing (TMED) proteins, also called p24 proteins, are members of a family of sorting receptors present in all representatives of the Eukarya and abundantly present in all subcompartments of the early secretory pathway, namely the endoplasmic reticulum (ER), the Golgi, and the intermediate compartment. Although essential during the bidirectional transport between the ER and the Golgi, there is still a lack of information regarding the TMED's structure across different subfamilies. Besides, although the presence of a TMED homo-oligomerization was suggested previously based on crystallographic contacts observed for the isolated Golgi Dynamics (GOLD) domain, no further analyses of its presence in solution were done. Here, we describe the first high-resolution structure of a TMED1 GOLD representative and its biophysical characterization in solution. The crystal structure showed a dimer formation that is also present in solution in a salt-dependent manner, suggesting that the GOLD domain can form homodimers in solution even in the absence of the TMED1 coiled-coil region. A molecular dynamics description of the dimer stabilization, with a phylogenetic analysis of the residues important for the oligomerization and a model for the orientation towards the lipid membrane, are also presented. (C) 2021 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. (AU)

• FAPESP's process: 15/16812-0 - Multi-User Equipment approved in grant 2014/15546-1: SEC-MALS
• Grantee: Richard Charles Garratt
• Support Opportunities: Multi-user Equipment Program
• 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
• 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