Phosphorylation of slit diaphragm proteins NEPHRIN and NEPH1 upon binding of HGF promotes podocyte repair

Autor: Bushra Rahman, Ehtesham Arif, Pei Wen, Zhe Han, Joshua H. Lipschutz, Avinash Singh, Sang Ho Kwon, Deepak Nihalani, Glenn P. Lobo, Pankaj Srivastava, Ashish K. Solanki, Wayne R. Fitzgibbon, Milos N. Budisavljevic, Christopher M. Furcht, Lawrence B. Holzman, Matthew J. Lazzara
Rok vydání: 2021
Předmět:
Kidney Glomerulus
Kidney
urologic and male genital diseases
Biochemistry
GST
glutathione S-transferase
Podocyte
Mice
HGF
PS
protamine sulfate
Phosphorylation
Receptor
MET
mesenchymal epithelial transition
biology
Hepatocyte Growth Factor
Podocytes
Chemistry
female genital diseases and pregnancy complications
Cell biology
phosphorylation–dephosphorylation
Intercellular Junctions
medicine.anatomical_structure
SHP-2
SH2 domain–containing protein tyrosine phosphatase-2
SHP-2
Slit diaphragm
Hepatocyte growth factor
Research Article
Glomerular Filtration Rate
Protein Binding
Signal Transduction
medicine.drug
Cell Line
FL
full length
Dephosphorylation
Nephrin
medicine
Animals
Humans
Molecular Biology
KD
knockdown
urogenital system
Membrane Proteins
Cell Biology
Actin cytoskeleton
ECD
extracellular domain
biology.protein
HGF
hepatocyte growth factor
Peptides
SNS
sticks-and-stones
CD
cytoplasmic domain
Zdroj: The Journal of Biological Chemistry
ISSN: 0021-9258
DOI: 10.1016/j.jbc.2021.101079
Popis: Phosphorylation (activation) and dephosphorylation (deactivation) of the slit diaphragm proteins NEPHRIN and NEPH1 are critical for maintaining the kidney epithelial podocyte actin cytoskeleton and, therefore, proper glomerular filtration. However, the mechanisms underlying these events remain largely unknown. Here we show that NEPHRIN and NEPH1 are novel receptor proteins for hepatocyte growth factor (HGF) and can be phosphorylated independently of the mesenchymal epithelial transition receptor in a ligand-dependent fashion through engagement of their extracellular domains by HGF. Furthermore, we demonstrate SH2 domain-containing protein tyrosine phosphatase-2-dependent dephosphorylation of these proteins. To establish HGF as a ligand, purified baculovirus-expressed NEPHRIN and NEPH1 recombinant proteins were used in surface plasma resonance binding experiments. We report high-affinity interactions of NEPHRIN and NEPH1 with HGF, although NEPHRIN binding was 20-fold higher than that of NEPH1. In addition, using molecular modeling we constructed peptides that were used to map specific HGF-binding regions in the extracellular domains of NEPHRIN and NEPH1. Finally, using an in vitro model of cultured podocytes and an ex vivo model of Drosophila nephrocytes, as well as chemically induced injury models, we demonstrated that HGF-induced phosphorylation of NEPHRIN and NEPH1 is centrally involved in podocyte repair. Taken together, this is the first study demonstrating a receptor-based function for NEPHRIN and NEPH1. This has important biological and clinical implications for the repair of injured podocytes and the maintenance of podocyte integrity.
Databáze: OpenAIRE
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