Identification of an iron-regulated 37,000-dalton protein in the cell envelope of Neisseria gonorrhoeae.

We examined the outer membrane proteins which appear during the growth of Neisseria gonorrhoeae F62 in complex medium supplemented with 25 microM Desferal mesylate, a potent iron chelator. Outer membranes were prepared by Sarkosyl extraction and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Several higher-molecular-weight (74,000 to greater than 94,000) proteins increased under iron-limiting conditions. In addition we observed the appearance of an iron-regulated protein with an apparent molecular weight of 37,000. This protein comigrated with the gonococcal protein I under normal Laemmli gel conditions. By increasing the ionic strength of the lower gel buffer, separation of protein I and the 37,000-dalton iron-regulated protein occurred. The 37,000-dalton protein stained poorly with Coomassie blue. However, when a silver stain was used, the protein appeared as a major component of the gonococcal outer membrane. Production of this 37,000-dalton protein was suppressed by the addition of iron to the medium. An iron-regulated protein with a similar molecular weight was observed in four clinical isolates and in an additional laboratory strain. Peptide mapping indicated that the 37,000-dalton protein was distinct from protein I and was identical between strains of the WI and WII serogroups.