Ligand‐induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity

Narayanan D, Larsen A, Gauger S, Adafia R, Hammershøi R, Hamborg L, Bruus‐Jensen J, Griem‐Krey N, Gee C, Frølund B, Stratton M, Kuriyan J, Kastrup J, Langkilde A, Wellendorph P, Solbak S, Protein Science 33(10) (2024) DOI

SASDU25 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) W403L hub domain with PIPA

Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L)
2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid
MWexperimental 208 kDa
MWexpected 185 kDa
VPorod 334 nm3
log I(s) 2.05×10-1 2.05×10-2 2.05×10-3 2.05×10-4
Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid small angle scattering data  s, nm-1
ln I(s)
Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid Guinier plot ln 2.06×10-1 Rg: 4.4 nm 0 (4.4 nm)-2 s2
(sRg)2I(s)/I(0)
Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid Kratky plot 1.104 0 √3 sRg
p(r)
Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid pair distance distribution function Rg: 4.4 nm 0 Dmax: 15 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) W403L hub domain with PIPA in 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH 6 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). One solute concentration of 6.80 mg/ml was measured at 20°C. 30 successive 0.095 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 data was collected for comparison with the CaMKIIa W403L hub domain without ligands present.

Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) (CaMKIIα W403L hub)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dodecamer
Mon. MW   15.4 kDa
 
UniProt   Q9UQM7 (345-475)
Sequence   FASTA
 
2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid (PIPA)
Mol. type   Other
Olig. state   Monomer
Mon. MW   0.3 kDa