Relative domain orientation of the L289K HIV ‐1 reverse transcriptase monomer

Xi Z, Ilina T, Guerrero M, Fan L, Sluis‐Cremer N, Wang Y, Ishima R, Protein Science 31(5) (2022) DOI

SASDNL3 – HIV-1 reverse transcriptase L289K mutation (extrapolated to zero concentration)

Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556)
MWI(0) 62 kDa
MWexpected 65 kDa
VPorod 90 nm3
log I(s) 1.31×10-1 1.31×10-2 1.31×10-3 1.31×10-4
Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) small angle scattering data  s, nm-1
ln I(s)
Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) Guinier plot ln 1.32×10-1 Rg: 3.2 nm 0 (3.2 nm)-2 s2
(sRg)2I(s)/I(0)
Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) Kratky plot 1.104 0 3 sRg
p(r)
Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) pair distance distribution function Rg: 3.2 nm 0 Dmax: 11.1 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) BUNCH model

Synchrotron SAXS data from solutions of HIV-1 reverse transcriptase (C38V/C280S, L289K) in 50 mM Bis-Tris pH 6.8, 100 mM NaCl, 1% glycerol, 0.02% w/v NaN3, pH 6.8 were collected on the 12ID-B SAXS/WAXS beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus 2M detector at a sample-detector distance of 1.9 m and at a wavelength of λ = 0.08856 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). 30 successive 1.500 second frames were collected at 20°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The resulting data is that calculated from the extrapolation of a concentration series SAXS data set to zero concentration.

Gag-Pol polyprotein (HIV-1 reverse transcriptase C38V/C280S, L289K truncated to 1 - 556) (p66_L289K)
Mol. type   Protein
Organism   Human immunodeficiency virus type 1 group M subtype B (isolate HXB2)
Olig. state   Monomer
Mon. MW   64.7 kDa
 
UniProt   P04585 (588-1143)
Sequence   FASTA