Western blot analysis of biotinylated proteins from P. aeruginosa PAO1 derivatives. (A) Lanes: 1, molecular masses of protein standards; 2 to 7, cell extracts from the PAO1 strain grown on glucose (lane 2), isoleucine (lane 3), leucine (lane 4), citronellol (lane 5), glucose plus leucine (lane 6), and glucose plus citronellol (lane 7). Proteins of 73, 70, 62, and 22 kDa identified with avidin-HRP conjugate, corresponding to biotinylated subunits from alpha subunits of geranyl-CoA carboxylase (AtuF), 3-methylcrotonyl-CoA carboxylase (LiuD), a putative acyl-CoA carboxylase, and acetyl-CoA carboxylase, respectively, are indicated. (B) Biotinylated proteins from cultures grown in glucose plus citronellol. Lanes: 1 and 3, PAO1 wild-type strain; 2, PAM liuD mutant; 4, PAO atuF mutant. The positions of the LiuD and AtuF proteins are indicated.