Crosstalk between regulatory elements in disordered TRPV4 N-terminus modulates lipid-dependent channel activity

Goretzki B, Wiedemann C, McCray B, Schäfer S, Jansen J, Tebbe F, Mitrovic S, Nöth J, Cabezudo A, Donohue J, Jeffries C, Steinchen W, Stengel F, Sumner C, Hummer G, Hellmich U, Nature Communications 14(1) (2023) DOI

SASDQF8 – G. gallus TRPV4 N-terminal Domain

Transient receptor potential cation channel subfamily V member 4
MWexperimental 43 kDa
MWexpected 43 kDa
VPorod 59 nm3
log I(s) 1.72×103 1.72×102 1.72×101 1.72×100
Transient receptor potential cation channel subfamily V member 4 small angle scattering data  s, nm-1
ln I(s)
Transient receptor potential cation channel subfamily V member 4 Guinier plot ln 1.72×103 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
Transient receptor potential cation channel subfamily V member 4 Kratky plot 1.104 0 √3 sRg
p(r)
Transient receptor potential cation channel subfamily V member 4 pair distance distribution function Rg: 4.1 nm 0 Dmax: 19 nm

Data validation

Fits and models

log I(s)
 s, nm-1
G. gallus TRPV4 N-terminal Domain Rg histogram Rg, nm
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Transient receptor potential cation channel subfamily V member 4 EOM/RANCH model
Synchrotron SAXS data from solutions of TRPV4 N-terminal domain in 20 mM Tris, 300 mM NaCl, 10 mM DTT, pH 7 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 40.00 μl sample at 10.0 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 2400 successive 0.250 second frames were collected through the entire SEC elution profile, where 107 frames were processed through the sample elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The molecule contains the isolated N-terminal domain from G. gallus TRPV4, spanning residues A2-D382. It consists of an intrinsically disordered region (A2-V134) and six Ankyrin repeats (F135-D382). The individual, unsubtracted SEC-SAXS data frames and additional modelling results are made available in the full entry zip archive.

Transient receptor potential cation channel subfamily V member 4 (TRPV4)
Mol. type   Protein
Organism   Gallus gallus
Olig. state   Monomer
Mon. MW   42.6 kDa
 
UniProt   A0A1D5PXA5 (2-382)
Sequence   FASTA