Skip to main content
. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Nat Chem Biol. 2019 Apr 1;15(5):463–471. doi: 10.1038/s41589-019-0251-4

Fig. 4. GacH and SccH modify their respective glycopolymers with sn-Gro-1-P.

Fig. 4.

a,b, Analysis of glycerol and phosphate content in GAC and SCC isolated from (a) GAS 5448 WT, ΔgacI, ΔgacH and ΔgacH complemented with native gacH or a catalytically-inactivated version of gacH (gacH-T530A), and (b) S. mutans WT, ΔsccH, and ΔsccH complemented with sccH or gacH. The concentration of phosphate and glycerol is presented relative to the WT strain. Bars and error bars represent the average and s.d., respectively (n=3 biologically independent samples). P-values were calculated and adjusted by 2-way ANOVA and Bonferroni’s multiple comparison test. c, DEAE-Sephacel elution profile of GAC isolated from ∼90 mg of GAS cell wall. Fractions were analyzed for carbohydrate (●) and phosphate (O). d–f, Identification of the enantiomeric form of GroP associated with GAC. d, The GroP isomers were recovered from GAC following alkaline hydrolysis and separated by liquid chromatography as outlined in Methods. The elution positions corresponding to standard Gro-2-P and sn-Gro-1-P/sn-Gro-3-P are indicated by the arrows. LC-MS analysis identifies two extracted ion chromatogram peaks for the molecular GroP ion m/z 171.004 [M-H]-, which eluted at (e) 9.48 and (f) 9.89 min. Based on the accurate mass and retention times, these two peaks were assigned as Gro-2-P and sn-Gro-1-P/sn-Gro-3-P respectively by comparison with authentic chemical standards. Experiments depicted in c–f were performed independently twice and yielded the same results.