FIG. 6.

Analysis of extracellular proteins from the wild type and the htrA mutant. (A) Supernatant proteins from overnight cultures were precipitated by 20% TCA, washed with acetone and resuspended in PBS. Equal amounts of proteins were loaded in each lane and samples were run on SDS-4 to 20% PAGE gels and stained with Coomassie blue. Bands corresponding to arrowheads were excised from stained gel and identified by mass spectrometry. Lanes: M, NEB prestained marker; 1, NG-8; 2, IBS101. Proteins identified by mass spectrometry are indicated at the right. (B) Western blot analysis of supernatant proteins separated by SDS-4 to 20% PAGE. The antibodies used are indicated below the gels. Lanes contain sample from NG-8 (lanes 1), IBS101 (lanes 2), and IBS101/pIB108 (lanes 3). Open arrows indicate different species of the reacted proteins.