Elution volume (ml)
12
14
16
18
20
Abs 295 nm
Abs 295 nm
Mw (kDa)
Mw (kDa)
0.0
0.2
0.4
0
20
40
60
80
0.02 mg/ml
0.08 mg/ml
0.22 mg/ml
0.02 mg/ml
0.08 mg/ml
0.22 mg/ml
66
29 kDa
Elution volume (ml)
12
14
16
18
20
22
0.0
0.5
1.0
1.5
0
30
60
90
120
0.02 mg/ml
0.14 mg/ml
0.21 mg/ml
0.02 mg/ml
0.14 mg/ml
0.21 mg/ml
220 156 kDa
a
b
c
[E.r. RT domain] mM
0.0
0.2
0.4
0.6
0.8
Mw (kDa)
0
30
60
90
120
150
Fig. 1 E.r. RT forms a dimer in solution in the absence or presence of D4A RNA.
Molecular weight distribution plot from SEC/MALS data for E.r. RT (panel a) and
E.r. RT:D4A RNA complex (panel b). Lines correspond to UV traces monitored at
295 nm (left axis); concentrations at the apex of the eluting peaks are listed in
the legend (in mg/ml); the M w are plotted as circles, or triangles (right y axis). For
clarity, only every tenth result of molecular weight measurement across the
eluting peak is plotted. Elution position of globular protein standards: betaamylase (220 kDa) and aldolase from rabbit muscles (156 kDa) are marked in
panel (b). Weight-average M w s determined from SEC/MALS analyses are plotted
as a function of the concentration at the apex of the eluting peak; filled circles for
E.r. RT-D4A RNA complex and open circles for E.r. RT protein alone (panel c) to
illustrate that E.r. RT forms a dimer that binds one D4A RNA per monomer