Human MICAL1: activation by the small GTPase Rab8 and small-angle X-ray scattering studies on the oligomerization state of MICAL1 and its complex with Rab8.

Esposito A, Ventura V, Petoukhov MV, Rai A, Svergun DI, Vanoni MA, Protein Sci (2018) Europe PMC

SASDDS9 – NADPH oxidase (H2O2 producing and [F-actin] oxidizing) MICAL1 (monomer) (Truncated MOCHLIM construct)

[F-actin]-monooxygenase MICAL1 (MoChLim)

MW^experimental	88	kDa
MW^expected	85	kDa
V^Porod	145	nm³

log I(s) 8.61×10³ 8.61×10² 8.61×10¹ 8.61×10⁰

[F-actin]-monooxygenase MICAL1 (MoChLim) small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

[F-actin]-monooxygenase MICAL1 (MoChLim) Guinier plot

ln 8.61×10³ R_g: 4 nm 0 (4 nm)^-2 s²

(sR_g)²I(s)/I(0)

[F-actin]-monooxygenase MICAL1 (MoChLim) Kratky plot

1.104 0 √3 sR_g

p(r)	R_g: 4.4 nm 0 D_max: 18 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.8

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.275
1.168

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

[F-actin]-monooxygenase MICAL1 (MoChLim) CORAL model

Synchrotron SAXS data, I(s) vs s (where s = 4πsinθ/λ, and 2θ is the scattering angle), were measured from the MoChLim construct of MICAL1 using continuous-flow size-exclusion chromatography SAXS (SEC-SAXS) on the EMBL-P12 beam line at the PETRA III storage ring (Hamburg, Germany). Data were collected a using a Pilatus 2M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm. The SEC mobile phase (0.5 ml/min) consisted of 20 mM Hepes/NaOH, pH 7.5, 50 mM NaCl, 2 mM MgCl2, 2 mM DTT, pH 7.5 (23°C). The sample load concentration onto the SEC column was 6.82 mg/ml (90 µl injection volume). The SAXS data measured from the SEC-elution (sample peak and buffer; 3600 successive 1 s frames) were normalized to the intensity of the transmitted beam and radially averaged. The scattering of an appropriate solvent-blank was subtracted from the sample frames to produce the scaled and averaged data displayed in this entry.

SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

[F-actin]-monooxygenase MICAL1 (MoChLim) (MoChLim)
Mol. type		Protein
Organism		Homo sapiens
Olig. state		Monomer
Mon. MW		85.2 kDa

UniProt		Q8TDZ2 (1-778)
Sequence		FASTA