FIG. 1.

Expression and activity of the PlsB26 mutant protein. (A) Membranes were prepared from strain SJ22 (plsB26) harboring either plasmid pRJ54 (plsB) or pJH3 (plsB26) or the empty vector (pACYC177), and the proteins were separated by sodium dodecyl sulfate-gel electrophoresis on 10% polyacrylamide gels. The samples were transferred to nitrocellulose membranes and immunoblotted with the M2 anti-Flag antibody to detect the expression of the epitope-tagged acyltransferases. (B) Membranes were prepared from strain 8 (wild type) (■) or strain SJ22 (plsB26) transformed with either the empty vector pACYC177 (□), pRJ54 (plsB) (●), or pJH3 (plsB26) (○). G3P acyltransferase assays were performed with the indicated amounts of membrane protein. The assays contained 0.1 M Tris-HCl (pH 8.6)–1-mg/ml bovine serum albumin–200 μM [¹⁴C]G3P (specific activity, 13 Ci/mmol)–50 μM palmitoyl-CoA and were incubated at 37°C for 15 min (12, 13).