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. 2017 Aug 29;292(42):17407–17417. doi: 10.1074/jbc.M117.804377

Figure 1.

Figure 1.

Isolation of ΔpimE suppressor mutants. A, PIMs/LM/LAM biosynthetic pathway. Mannosyltransferases involved in the biosynthesis of the LM intermediate are unknown. MptA is α1–6-mannosyltransferase involved in the mannan backbone elongation, whereas MptC is α1–2-mannosyltransferase that adds monomannose side chains. The mannose donors, GDP-mannose (GDP-Man) and PPM, are indicated for mannosyltransferase reactions. B, colony morphology of WT, ΔpimE, and the suppressor mutant S1. The suppressor mutant S1 (as well as the other 21 suppressor mutants; not shown) restored WT colony morphology. Scale bar, 1 cm. C, profile of PIMs purified from the suppressor mutants. Lipid extracts of WT, ΔpimE, and the suppressor mutants S1–S6 were separated by TLC and visualized by orcinol staining. Only a part of the TLC plate is shown. None of the suppressor mutants were able to restore the production of AcPIM6 (arrowhead). See supplemental Fig. S1 for the other suppressor mutants. D, LM/LAM profile of the suppressor mutants, showing the small LM/LAM phenotype of the mutant S1. Extracts of LM/LAM were separated on SDS-PAGE and visualized by ProQ Emerald 488 glycan staining. See supplemental Fig. S2 for the other suppressor mutants.