The outer membrane of Gram-negative bacteria contains a diverse range of proteins that typically adopt a β-barrel fold. αβ-Barrels, which are often misannotated simply as “β-barrels”, are thought to follow the “standard” β-barrel folding trajectory, but this hypothesis does not account for their large α-barrel domain that extends the β-barrel pore into the periplasm. While the presence of both an α-barrel and β-barrel domain would suggest this class of proteins requires two distinct folding steps, many (but not all) αβ-barrels are N-terminally acylated (i.e. lipoproteins) and inherently follow an alternative biogenesis route compared to their non-lipoprotein counterparts. By taking an RNAseq approach, cells that were engineered to be deficient in an essential component of their folding pathway had an increased reliance on the more elusive components of the αβ-barrel biogenesis machinery. An exploration of the chaperones that were identified now paints a more accurate picture of the unique folding pathway of complex substrates, and how they navigate the protein and lipoprotein folding interface.