The role of linker length and composition in actin binding and bundling by palladin.

Sargent RA, Bradford CW, Hughes LM, Ta NH, Limpiado MJ, Vattepu R, Beck MR, Biochem J 483(3):301-318 (2026) Europe PMC

SASDX77 – Palladin Ig3-Ig4 tandem domain with linker swapped for the shorter Ig4-Ig5 Linnker

Palladin Ig3-45Linker_Ig4
MWI(0) 29 kDa
MWexpected 22 kDa
VPorod 27 nm3
log I(s) 7.85×10-3 7.85×10-4 7.85×10-5 7.85×10-6
Palladin Ig3-45Linker_Ig4 small angle scattering data  s, nm-1
ln I(s)
Palladin Ig3-45Linker_Ig4 Guinier plot ln 7.86×10-3 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
Palladin Ig3-45Linker_Ig4 Kratky plot 1.104 0 3 sRg
p(r)
Palladin Ig3-45Linker_Ig4 pair distance distribution function Rg: 3 nm 0 Dmax: 10.4 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Palladin Ig3-Ig4 tandem domain with linker swapped for the shorter Ig4-Ig5 Linnker Rg histogram Rg, nm
Palladin Ig3-45Linker_Ig4 EOM/RANCH model
Palladin Ig3-45Linker_Ig4 EOM/RANCH model
Palladin Ig3-45Linker_Ig4 EOM/RANCH model

Synchrotron SAXS data from solutions of Palladin Ig3-Ig4 tandem domain with linker swapped for the shorter Ig4-Ig5 Linnker in HEPES, pH 7.4 were collected on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSRL) storage ring (Menlo Park, CA, USA) using a Rayonix MX225-HE detector (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). . The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Synchrotron SAXS data from solutions of the Ig-like C2-type 3 and Ig-like C2-type 4 domains (Ig34) of palladin where the wildtype linker region has been replaced by shorter linker found between Ig4 and Ig5 domains in 20 mM HEPES pH 7.4, 1 mM DTT, 100 mM NaCl were collected on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSR; Menlo Park, CA, USA) using a Rayonix MX225-HE detector at a sample-detector distance of 1.2 m and at a wavelength of λ = 0.113 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.5 and 10.0 mg/ml were measured at 20°C. 12 successive 1 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Palladin Ig3-45Linker_Ig4 (Palladin Ig3-45L-Ig4)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   22.3 kDa
 
UniProt   Q9ET54 (1022-1257)
Sequence   FASTA