Asparagine-84, a regulatory allosteric site residue, helps maintain the quaternary structure of Campylobacter jejuni dihydrodipicolinate synthase.

Majdi Yazdi M, Saran S, Mrozowich T, Lehnert C, Patel TR, Sanders DAR, Palmer DRJ, J Struct Biol :107409 (2019) Europe PMC

SASDG36 – 4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant)

4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant)
MWexperimental 78 kDa
MWexpected 65 kDa
VPorod 108 nm3
log I(s) 7.70×10-2 7.70×10-3 7.70×10-4 7.70×10-5
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) small angle scattering data  s, nm-1
ln I(s)
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) Guinier plot ln 7.70×10-2 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) Kratky plot 1.104 0 √3 sRg
p(r)
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) pair distance distribution function Rg: 3.1 nm 0 Dmax: 9.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) DAMFILT model
4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) DAMMIF model
Synchrotron SAXS data from solutions of N84D mutant 4-hydroxy-tetrahydrodipicolinate synthase in 20 mM Tris-HCl, 150 mM NaCl, pH 8 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 = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 10 mg/ml was injected at a 0.16 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column . 619 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.

The models displayed in this entry represent: Top, the averaged (DAMFILT) model obtained from the spatial alignment of several individual DAMMIF models (volume and bead occupancy corrected) and; Bottom, and individual DAMMIF model representative and corresponding fit. Additional individual DAMMIF models (in P1 symmetry) are provided in the full entry zip archive.

4-hydroxy-tetrahydrodipicolinate synthase (N84D mutant) (HTPA synthase N84D)
Mol. type   Protein
Organism   Campylobacter jejuni
Olig. state   Dimer
Mon. MW   32.7 kDa
 
UniProt   Q9PPB4 (1-298)
Sequence   FASTA