Lp. Yu et al., BACKBONE DYNAMICS OF THE C-TERMINAL DOMAIN OF ESCHERICHIA-COLI TOPOISOMERASE-I IN THE ABSENCE AND PRESENCE OF SINGLE-STRANDED-DNA, Biochemistry, 35(30), 1996, pp. 9661-9666
The backbone dynamics of the C-terminal DNA-binding domain of Escheric
hia coli topoisomerase I has been characterized in the absence and pre
sence of single-stranded DNA by NMR spectroscopy. N-15 spin-lattice re
laxation times (T-1), spin-spin relaxation times (T-2), and heteronucl
ear NOEs were determined for the uniformly N-15-labeled protein. These
data were analyzed by using the model-free formalism to derive the mo
del-free parameters (S-2, tau(e), and R(ex)) for each backbone N-H bon
d vector and the overall molecular rotational correlation time (tau(m)
), The molecular rotational correlation time tau(m) was determined to
be 7.49 +/- 0.36 ns for the free and 12.7 +/- 1.07 ns for the complexe
d protein. Several residues were found to be much more mobile than the
average, including 11 residues at the N-terminus, 2 residues at the C
-terminus, and residues 25 and 31-35 which are located in a region of
the protein that binds to DNA. The binding of ssDNA to the free protei
n causes a slight increase in the order parameters (S-2) for a small n
umber of residues and a slight decrease in the order parameters (S-2)
for the majority of the residues. In particular, upon binding to ssDNA
, the mobility of the first alpha-helix and the two beta-sheets was sl
ightly increased, and the mobility of a few specific residues in the l
oops/turns was restricted. These results differ from the previous stud
ies on the backbone dynamics of molecular complexes in which reduced m
obilities were typically observed upon ligand binding.