A staphylococcal anti-sigma factor possesses a single-domain, carries different denaturant-sensitive regions and unfolds via two intermediates
Fig 2
(A) The sequence of rRsbWwith the cleavage sites of chymotrypsin (Ch), proteinase K (Pr), and trypsin (Tr). All of the predicted cut sites, determined using a computational tool [31], are presented by verticle lines on both sides oftheamino acid sequence. The experimentally-detected cut sites (I-VI) are shown by downward arrows over the rRsbW sequence. The polyhistidine tag carrying regionofrRsbW is composed of amino acid residues 1 to 36. (B, C, D) Analyses of the rRsbW fragments by SDS-13.5% PAGE after Ch, Pr, and Tr digestions, respectively. Arrowhead and I−VI denote the intact rRsbW and its major proteolytic fragments of rRsbW, respectively. (E, F, G) Western blotting analyses of the Ch -, Pr -, and Tr -digested rRsbW fragments using an anti-his antibody. The molecular masses (in kDa) of marker proteins are shown at the right side of the gel/blot pictures.