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Figure 1.
Fig. 1. Identification and characterisation of new stable
PspF[1–275] WT•σ^54 or Eσ^54 complexes in the presence of
AMP–AlF[x]. (a) Native gel showing the complexes formed by
PspF[1–275] WT (5 μM) with or without σ^54 (1 μM) and with
or without RNA polymerase (0.15 μM) in the presence of AlF[x]
reactant [AlC[3] (0.4 mM) + NaF (5 mM)] and different
nucleotides (4 mM, as indicated). The sample was loaded onto
native 4.5% PAGE and proteins were detected by Coomassie blue
staining. ANP indicates AMP or ADP. (b) Gel filtration profiles
of samples containing PspF[1–275] WT (64 μM) with or without
σ^54 (30 μM) and with or without AMP–AlF[x] or ADP–AlF[x]
(as indicated) chromatographed through a Superdex 200 column (10
mm × 300 mm, 24 ml, GE Healthcare) at 4 °C. The scale
bars give the scale of the ordinate axis; absorption units (AU)
correspond to an A[280 nm] of 1.
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