Order from disorder in the sarcomere: FATZ forms a fuzzy but tight complex and phase-separated condensates with α-actinin.

Sponga A, Arolas JL, Schwarz TC, Jeffries CM, Rodriguez Chamorro A, Kostan J, Ghisleni A, Drepper F, Polyansky A, De Almeida Ribeiro E, Pedron M, Zawadzka-Kazimierczuk A, Mlynek G, Peterbauer T, Doto P, Schreiner C, Hollerl E, Mateos B, Geist L, Faulkner G, Kozminski W, Svergun DI, Warscheid B, Zagrovic B, Gautel M, Konrat R, Djinović-Carugo K, Sci Adv 7(22) (2021) Europe PMC

SASDJP6 – Sarcomeric fuzzy α-actinin-2/FATZ-1 complex (hd-α-actinin-2/Δ91-FATZ-1)

Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2)
Half dimer of α-actinin-2
MWexperimental 147 kDa
MWexpected 129 kDa
VPorod 242 nm3
log I(s) 1.40×104 1.40×103 1.40×102 1.40×101
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 small angle scattering data  s, nm-1
ln I(s)
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 Guinier plot ln 1.41×104 Rg: 5.8 nm 0 (5.8 nm)-2 s2
(sRg)2I(s)/I(0)
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 Kratky plot 1.104 0 3 sRg
p(r)
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 pair distance distribution function Rg: 6.1 nm 0 Dmax: 22.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Sarcomeric fuzzy α-actinin-2/FATZ-1 complex (hd-α-actinin-2/Δ91-FATZ-1) Rg histogram Rg, nm
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 EOM/RANCH model
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 EOM/RANCH model
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 EOM/RANCH model
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 EOM/RANCH model
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 EOM/RANCH model

log I(s)
 s, nm-1
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 CORAL model

log I(s)
 s, nm-1
Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) Half dimer of α-actinin-2 CORAL model

Synchrotron SAXS data from solutions the hd-α-actinin-2/Δ91-FATZ-1 complex in 50 mM Tris-HCl 150 mM NaCl, pH 7.5 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 100.00 μl sample at 5 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superose 6 Increase 10/300 column at 20°C. 55 successive 0.995 second frames were collected through the major SEC-elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Δ91 construct of FATZ-1 (alias myozenin-1 or calsarcin-2) (Δ91-FATZ-1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   21.5 kDa
 
UniProt   Q9NP98 (92-299)
Sequence   FASTA
 
Half dimer of α-actinin-2 (hd-α-actinin-2)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   107.0 kDa
 
UniProt   P35609 (1-894)
Sequence   FASTA