Crystal Structure of the N-Terminal RNA Recognition Motif of mRNA Decay Regulator AUF1
Preparation of the AUF1- protein. (a) Domain organization of AUF1-p37 and truncated constructs. (b) Chromatogram of size-exclusion chromatography for AUF1-. The dotted line indicates a chromatogram in the absence of RNaseA. The black line represents the peak chromatogram in the presence of RNaseA. The standard molecular masses for SEC experiments were obtained from the following proteins: β-amylase, 200 kDa; albumin, 66 kDa; carbonic anhydrase, 29 kDa; and cytochrome C, 12.4 kDa. (c) Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the size-exclusion chromatographic fractions, as shown in (b). The labels above the gel indicate: M, low-range marker (Bio-Rad); 1, whole-cell fraction; 2, soluble fraction; 3, insoluble fraction; 4, flow-through fraction; 5, wash fraction with 25 mM imidazole; 6, elution fraction with 250 mM imidazole; 7–14, fractions from the size-exclusion chromatography.
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