Varying selection pressure for a Na+ sensing site in epithelial Na+ channel subunits reflect divergent roles in Na+ homeostasis DOI Creative Commons
Xue‐Ping Wang, Priyanka Srinivasan, Mustapha El Hamdaoui

et al.

Molecular Biology and Evolution, Journal Year: 2024, Volume and Issue: 41(8)

Published: Aug. 1, 2024

The epithelial Na+ channel (ENaC) emerged early in vertebrates and has played a role fluid homeostasis throughout vertebrate evolution. We previously showed that proteolytic activation of the evolved at water-to-land transition vertebrates. Sensitivity to extracellular Na+, known as self-inhibition, reduces ENaC function when concentrations are high is distinctive feature channel. A fourth subunit, δ, jawed fishes from an α subunit gene duplication. Here, we analyzed 849 δ sequences found key Asp postulated binding site was nearly always present but frequently lost (e.g. human). Analysis evolution codon substitution rates provide evidence ancestral had purifying selection for relaxed after its divergence coinciding with loss expression renal tissues. also show proposed bona fide by conferring novel channels comprising human subunits. Together, our findings self-inhibition improves fitness through

Language: Английский

Role of Actin Dynamics in the Control of Proteolytically Induced ENaC-Like Channel Activity in Human Leukemia Cells DOI
Daria V. Lysikova, Valeria Y. Vasileva, Olga Tsaplina

et al.

Cell and Tissue Biology, Journal Year: 2025, Volume and Issue: 19(1), P. 48 - 54

Published: Jan. 20, 2025

Language: Английский

Citations

0
Automated patch-clamp recordings for detecting activators and inhibitors of the epithelial sodium channel (ENaC) DOI Creative Commons
Florian Sure, Markus Rapedius,

Alexei Diakov

et al.

Pflügers Archiv - European Journal of Physiology, Journal Year: 2025, Volume and Issue: unknown

Published: May 8, 2025

Abstract The epithelial sodium channel (ENaC) is crucial for absorption in several tissues including lung and kidney. Its involvement various renal pulmonary disorders makes ENaC a potential drug target. High-throughput screening using the automated patch-clamp (APC) technique appears to be promising approach discover novel modulators with (patho-)physiological therapeutic implications. aim of this methodological study was establish APC measurements ENaC-mediated currents. First, we confirmed functional expression HEK293 cell line stably transfected human αβγ-ENaC conventional manual whole-cell recordings. For measurements, standard enzymatic cell-detachment procedure used prepare single suspensions. This resulted high success rate recordings amiloride inhibitable Using γ-inhibitory peptide small molecule activator S3969, demonstrate that could reveal inhibitory as well stimulatory effects on ENaC. Interestingly, protocol partial proteolytic activation. portion proteolytically activated channels reduced by prolonged incubation suspended cells culture medium. recovery enhanced relative effect chymotrypsin, prototypical serine protease known cause Thus, may particularly useful identifying activators mimicking In conclusion, have established high-throughput method identification inhibitors APC.

Language: Английский

Citations

0
Influence of proteolytic cleavage of ENaC’s γ subunit upon Na+ and K+ handling DOI
Evan C. Ray, Andrew J. Nickerson, Shaohu Sheng

et al.

AJP Renal Physiology, Journal Year: 2024, Volume and Issue: 326(6), P. F1066 - F1077

Published: April 18, 2024

The epithelial Na

Language: Английский

Citations

3
Loss of the alpha subunit distal furin cleavage site blunts ENaC activation following Na+ restriction DOI Creative Commons
Andrew J. Nickerson, Shaohu Sheng,

Natalie A. Cox

et al.

The Journal of Physiology, Journal Year: 2024, Volume and Issue: 602(17), P. 4309 - 4326

Published: Aug. 28, 2024

Abstract Epithelial Na + channels (ENaCs) are activated by proteolysis of the α and γ subunits at specific sites flanking embedded inhibitory tracts. To examine role subunit in channel activation vivo , we generated mice lacking distal furin cleavage site (α F2M mice). On a normal control diet, no differences ENaC protein abundance kidney or colon were noted between wild‐type (WT) mice. Patch‐clamp analyses revealed similar levels activity tubules, while physiologically relevant blood chemistry aldosterone detected. Male did exhibit diminished colon, as measured amiloride‐sensitive short‐circuit current ( I SC ). Following dietary restriction, WT had natriuretic colonic responses to amiloride. However, single‐channel was significantly lower tubules from ‐restricted compared with littermates. expression also enhanced vs . mice, association higher levels. These data provide evidence that disrupting impairs requiring compensation response restriction. image Key points The epithelial (ENaC) is proteolytic vitro but key questions regarding terms whole‐animal physiology remain be addressed. We studied importance this mechanism generating mouse model genetic disruption ENaC's found not phenotype under conditions, have impaired (channel open probability) during salt function organ level preserved salt‐restricted associated increased subunits, suggesting required maintain homeostasis. results first regulator

Language: Английский

Citations

1
Going with the flow: New insights regarding flow induced K+ secretion in the distal nephron DOI Creative Commons
Samia Lasaad, Andrew J. Nickerson, Gilles Crambert

et al.

Physiological Reports, Journal Year: 2024, Volume and Issue: 12(20)

Published: Oct. 1, 2024

Abstract K + secretion in the distal nephron has a critical role homeostasis and is primary route by which lost from body. Renal enhanced increases dietary intake tubular flow rate nephron. This review addresses new important insights regarding mechanisms underlying flow‐induced (FIKS). While basal mediated renal outer medullary (ROMK) channels principal cells (PCs), FIKS large conductance, Ca 2+ /stretch activated (BK) intercalated (ICs), distinct cell type. BK channel activation requires an increase intracellular concentration ([Ca ] i ), both PCs ICs exhibit [Ca response to fluid rate, associated with diameter. PIEZO1, mechanosensitive, nonselective cation channel, expressed basolateral membranes of ICs, where it functions as mechanosensor. The loss transients channel‐mediated microperfused collecting ducts isolated mice IC‐specific deletion Piezo1 CCD underscores importance PIEZO1 regulation transport.

Language: Английский

Citations

1
Varying selection pressure for a Na+sensing site in epithelial Na+channel subunits reflect divergent roles in Na+homeostasis DOI Creative Commons
Xue‐Ping Wang, Priyanka Srinivasan, Mustapha El Hamdaoui

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 21, 2024

Abstract The epithelial Na + channel (ENaC) emerged early in vertebrates and has played a role fluid homeostasis throughout vertebrate evolution. We previously showed that proteolytic activation of the evolved at water to land transition vertebrates. Sensitivity extracellular , known as self-inhibition, reduces ENaC function when concentrations are high is distinctive feature channel. A fourth subunit, δ, jawed fishes from an α subunit gene duplication. Here, we analyzed 846 δ sequences found key Asp postulated binding site was nearly always present but frequently lost (e.g., human). Analysis evolution codon substitution rates provide evidence ancestral had purifying selection for relaxed after its divergence coinciding with loss expression renal tissues. also proposed bona fide by conferring novel channels comprising human subunits. Together, our findings self-inhibition improves fitness through

Language: Английский

Citations

0
Varying selection pressure for a Na+ sensing site in epithelial Na+ channel subunits reflect divergent roles in Na+ homeostasis DOI Creative Commons
Xue‐Ping Wang, Priyanka Srinivasan, Mustapha El Hamdaoui

et al.

Molecular Biology and Evolution, Journal Year: 2024, Volume and Issue: 41(8)

Published: Aug. 1, 2024

The epithelial Na+ channel (ENaC) emerged early in vertebrates and has played a role fluid homeostasis throughout vertebrate evolution. We previously showed that proteolytic activation of the evolved at water-to-land transition vertebrates. Sensitivity to extracellular Na+, known as self-inhibition, reduces ENaC function when concentrations are high is distinctive feature channel. A fourth subunit, δ, jawed fishes from an α subunit gene duplication. Here, we analyzed 849 δ sequences found key Asp postulated binding site was nearly always present but frequently lost (e.g. human). Analysis evolution codon substitution rates provide evidence ancestral had purifying selection for relaxed after its divergence coinciding with loss expression renal tissues. also show proposed bona fide by conferring novel channels comprising human subunits. Together, our findings self-inhibition improves fitness through

Language: Английский

Citations

0