Evidence That Intracellular Stages of Leishmania major Utilize Amino Sugars as a Major Carbon Source
Figure 2
GND is required for growth on GlcN or GlcNAc.
(A) Wild type (WT), Δgnd and complemented strains (Δgnd +GND, Δgnd +GNDΔSKL were suspended in CDM with or without Glc, GlcN or GlcNAc (13 mM). The optical density of cultures (OD600) at day 4 are shown. (B) L. major wild type, Δgnd, Δgnd +GND and Δgnd +GNDΔSKL promastigotes were lysed and GND activity determined by measuring production of hexose-6-phosphates from GlcN6P. (C,D) Wild type, Δgnd and complemented strains were labeled with 3H-GlcN and incorporation of 3H-label into (C) lipids and (D) the carbohydrate reserve polymer mannogen, assessed by HPTLC and fluorography. Major lipid species comprise PtdEtn, PtdCho, PtdIno, inositolphosphoceramide (IPC), PtdInoP and GPI species (asterix); Mannose (M) and mannogen oligomers (Mn).