FIG. 8.

Radiolabeling of GldH-His, GldB-His, and GldD with [³H]palmitate. Cells of F. johnsoniae were labeled with [³H]palmitate or [³H]glutamate. Proteins were isolated by precipitation with Ni-NTA His-Bind resin (A) or by immunoprecipitation (B), separated by SDS-PAGE, and detected by autoradiography. (A) Labeling of His-tagged Gld proteins. Lane 1, gldH mutant CJ1043 expressing GldH-His from pTB41, labeled with [³H]palmitate. Lane 2, gldB mutant CJ569 expressing GldB-His from pTB39, labeled with [³H]palmitate. Lane 3, CJ1043 expressing GldH-His from pTB41, labeled with [³H]glutamate. Lane 4, CJ569 expressing GldB-His from pTB39, labeled with [³H]glutamate. (B) Labeling of GldD. Lane 1, Wild-type F. johnsoniae labeled with [³H]palmitate. Lane 2, gldD mutant CJ282 labeled with [³H]palmitate. Lane 3, CJ282 complemented with pMM213, labeled with [³H]palmitate. Lane 4, wild-type F. johnsoniae labeled with [³H]glutamate. Lane 5, gldD mutant CJ282 labeled with [³H]glutamate. Lane 6, CJ282 complemented with pMM213, labeled with [³H]glutamate.