Figure 1.

Purification and tRNA-Processing Activity of Recombinant T. brucei PRORP1 and PRORP2

(A) SDS-PAGE of recombinant PRORP1 and PRORP2 purified by affinity chromatography (AC), or affinity chromatography with subsequent size exclusion chromatography (SEC) is shown; molecular weight of marker proteins indicated in kDa.

(B) Size exclusion chromatography profile of purified recombinant PRORP1 (blue) and PRORP2 (red) is illustrated. The peak at about 8 ml represents aggregated protein eluting in the void volume. The peak positions of molecular weight marker proteins resolved under identical conditions are indicated.

(C) RNase P activity of recombinant PRORP1 and PRORP2 is presented. A T. brucei tRNA_i^Met precursor was incubated with PRORP1, PRORP2, or E. coli RNase P. 5′ end-labeled substrate RNA and cleavage product (indicated by icons to the right) were resolved by denaturing PAGE.

(D) Same as (C), but T. brucei tRNA^Phe precursor used as substrate is shown.

(E) Same as (C), but T. brucei tRNA^His precursor used as substrate is presented.

(F) Same as (C), but E. coli tRNA^Tyrsu₃⁺ precursor used as substrate is illustrated.

(G) Same as (C), but tRNA^Gly precursor from Thermus thermophilus used as substrate is demonstrated.

See Figure S1 for alignment of trypanosomatid PRORP sequences to human and plant PRORP.