Arabidopsis and Chlamydomonas phosphoribulokinase crystal structures complete the redox structural proteome of the Calvin-Benson cycle.

Gurrieri L, Del Giudice A, Demitri N, Falini G, Pavel NV, Zaffagnini M, Polentarutti M, Crozet P, Marchand CH, Henri J, Trost P, Lemaire SD, Sparla F, Fermani S, Proc Natl Acad Sci U S A 116(16):8048-8053 (2019) Europe PMC

SASDDH9 – Chloroplastic phosphoribulokinase (collected using SEC-SAXS)

Phosphoribulokinase, chloroplastic
MWI(0) 70 kDa
MWexpected 78 kDa
VPorod 115 nm3
log I(s) 4.20×101 4.20×100 4.20×10-1 4.20×10-2
Phosphoribulokinase, chloroplastic small angle scattering data  s, nm-1
ln I(s)
Phosphoribulokinase, chloroplastic Guinier plot ln 4.20×101 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
Phosphoribulokinase, chloroplastic Kratky plot 1.104 0 √3 sRg
p(r)
Phosphoribulokinase, chloroplastic pair distance distribution function Rg: 3.5 nm 0 Dmax: 11.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Phosphoribulokinase, chloroplastic PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of chloroplastic phosphoribulokinase (collected using SEC-SAXS) in Tris-HCl 50 mM 150 mM KCl, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 4°C. 80 successive 1 second frames were collected through the SEC-elution profile. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of an appropriate solvent-blank (eluate SEC-buffer) was subtracted.

Additional SEC parameters: Column type: Superdex 200 10/300 GL (GE Healthcare); Flow rate: 0.5 ml/min; Sample injection concentration: 6.1 mg/ml; Injection volume: 100 µl.

Phosphoribulokinase, chloroplastic (CrPRK)
Mol. type   Protein
Organism   Chlamydomonas reinhardtii
Olig. state   Dimer
Mon. MW   38.9 kDa
 
UniProt   P19824 (32-375)
Sequence   FASTA
 
PDB ID   6H7G