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

SASDU45 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M)

MW^experimental	193	kDa
MW^expected	217	kDa
V^Porod	329	nm³

log I(s) 1.51×10^-1 1.51×10^-2 1.51×10^-3 1.51×10^-4

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) Guinier plot

ln 1.52×10^-1 R_g: 4.7 nm 0 (4.7 nm)^-2 s²

(sR_g)²I(s)/I(0)

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) Kratky plot

1.104 0 √3 sR_g

p(r)	R_g: 4.8 nm 0 D_max: 18 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.5

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.244
1.001

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) PYMOL model

Synchrotron SAXS data from solutions of calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain in 20 mM MES, 150 mM NaCl, 1 mM DTT, 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 3.40 mg/ml was measured at 20°C. 40 successive 0.090 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.

Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) (CaMKIIα 6x hub)
Mol. type		Protein
Organism		Homo sapiens
Olig. state		14-mer
Mon. MW		15.5 kDa

UniProt		Q9UQM7 (345-475)
Sequence		FASTA

PDB ID		6OF8