FIGURE 4.

Glycosylation of protein substrates in glycocompetent E. coli. Representative Western blot analysis of periplasmically-localized AcrA (A) and PEB3 (C) translocated via the Sec or TAT pathway in E. coli (n = 3). Total AcrA or PEB3 was detected with anti-His antibody, and N-linked glycosylated proteins were detected with anti-N-glycan antibody (R1). Non-glycosylated (g₀), mono-glycosylated (g₁), and di-glycosylated (g₂) species are indicated. Average glycosylation of AcrA (B) and PEB3 (D) from three biological replicates was determined by densitometry analysis of non-glycosylated and glycosylated protein bands (*, p < 0.02; **, p < 0.002).