Transglutaminase-Mediated Cross-Linking of Tropoelastin to Fibrillin Stabilises the Elastin Precursor Prior to Elastic Fibre Assembly.

Lockhart-Cairns MP, Newandee H, Thomson J, Weiss AS, Baldock C, Tarakanova A, J Mol Biol 432(21):5736-5751 (2020) Europe PMC

SASDNW3 – Fibrillin-1 fragment PF2 crosslinked to tropoelastin by transglutaminase 2

Fibrillin-1
Elastin
MWexperimental 155 kDa
MWexpected 104 kDa
log I(s) 1.77×10-1 1.77×10-2 1.77×10-3 1.77×10-4
Fibrillin-1 Elastin small angle scattering data  s, nm-1
ln I(s)
Fibrillin-1 Elastin Guinier plot ln 1.77×10-1 Rg: 7.1 nm 0 (7.1 nm)-2 s2
(sRg)2I(s)/I(0)
Fibrillin-1 Elastin Kratky plot 1.104 0 √3 sRg
p(r)
Fibrillin-1 Elastin pair distance distribution function Rg: 7.1 nm 0 Dmax: 23.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Fibrillin-1 Elastin DAMMIF model
Synchrotron SAXS data from solutions of fibrillin-1 fragment PF2 crosslinked to tropoelastin by transglutaminase 2 in Tris buffered saline, pH 7.4 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 4 m and at a wavelength of λ = 0.099 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 1.00 mg/ml was measured at 25°C. 10 successive 3 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.

Fibrillin-1
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   44.2 kDa
 
UniProt   P35555 (330-722)
Sequence   FASTA
 
Elastin
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   60.0 kDa
 
UniProt   P15502 (27-724)
Sequence   FASTA