Phase Separation of MAGI2-Mediated Complex Underlies Formation of Slit Diaphragm Complex in Glomerular Filtration Barrier.
Adaptor Proteins, Signal Transducing
/ chemistry
Animals
Biophysical Phenomena
Cell Adhesion Molecules
/ genetics
Cytoskeletal Proteins
/ chemistry
Fluorescence Recovery After Photobleaching
Glomerular Filtration Barrier
/ chemistry
Green Fluorescent Proteins
Guanylate Kinases
/ chemistry
Humans
Membrane Proteins
/ chemistry
Mice
Molecular Structure
Mutation
Nerve Tissue Proteins
/ chemistry
Phase Transition
Podocytes
/ metabolism
Protein Interaction Domains and Motifs
MAGI2
cell adhesion
cytoskeleton
nephrin
nephrotic syndrome
phase separation
slit diaphragm
Journal
Journal of the American Society of Nephrology : JASN
ISSN: 1533-3450
Titre abrégé: J Am Soc Nephrol
Pays: United States
ID NLM: 9013836
Informations de publication
Date de publication:
08 2021
08 2021
Historique:
received:
12
11
2020
accepted:
22
03
2021
entrez:
31
7
2021
pubmed:
1
8
2021
medline:
21
10
2021
Statut:
ppublish
Résumé
Slit diaphragm is a specialized adhesion junction between the opposing podocytes, establishing the final filtration barrier to urinary protein loss. At the cytoplasmic insertion site of each slit diaphragm there is an electron-dense and protein-rich cellular compartment that is essential for slit diaphragm integrity and signal transduction. Mutations in genes that encode components of this membrane-less compartment have been associated with glomerular diseases. However, the molecular mechanism governing formation of compartmentalized slit diaphragm assembly remains elusive. We systematically investigated the interactions between key components at slit diaphragm, such as MAGI2, Dendrin, and CD2AP, through a combination of biochemical, biophysical, and cell biologic approaches. We demonstrated that MAGI2, a unique MAGUK family scaffold protein at slit diaphragm, can autonomously undergo liquid-liquid phase separation. Multivalent interactions among the MAGI2-Dendrin-CD2AP complex drive the formation of the highly dense slit diaphragm condensates at physiologic conditions. The reconstituted slit diaphragm condensates can effectively recruit Nephrin. A nephrotic syndrome-associated mutation of Key components at slit diaphragm (
Sections du résumé
BACKGROUND
Slit diaphragm is a specialized adhesion junction between the opposing podocytes, establishing the final filtration barrier to urinary protein loss. At the cytoplasmic insertion site of each slit diaphragm there is an electron-dense and protein-rich cellular compartment that is essential for slit diaphragm integrity and signal transduction. Mutations in genes that encode components of this membrane-less compartment have been associated with glomerular diseases. However, the molecular mechanism governing formation of compartmentalized slit diaphragm assembly remains elusive.
METHODS
We systematically investigated the interactions between key components at slit diaphragm, such as MAGI2, Dendrin, and CD2AP, through a combination of biochemical, biophysical, and cell biologic approaches.
RESULTS
We demonstrated that MAGI2, a unique MAGUK family scaffold protein at slit diaphragm, can autonomously undergo liquid-liquid phase separation. Multivalent interactions among the MAGI2-Dendrin-CD2AP complex drive the formation of the highly dense slit diaphragm condensates at physiologic conditions. The reconstituted slit diaphragm condensates can effectively recruit Nephrin. A nephrotic syndrome-associated mutation of
CONCLUSIONS
Key components at slit diaphragm (
Identifiants
pubmed: 34330769
pii: 00001751-202108000-00017
doi: 10.1681/ASN.2020111590
pmc: PMC8455267
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
CD2-associated protein
0
Cell Adhesion Molecules
0
Cytoskeletal Proteins
0
MAGI3 protein, human
0
Membrane Proteins
0
Nerve Tissue Proteins
0
dendrin
0
nephrin
0
Green Fluorescent Proteins
147336-22-9
Guanylate Kinases
EC 2.7.4.8
MAGI2 protein, human
EC 2.7.4.8
Magi1 protein, mouse
EC 2.7.4.8
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1946-1960Informations de copyright
Copyright © 2021 by the American Society of Nephrology.
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