Figure 3. PM20D1 regulates the levels of N-acyl amino acids in vivo.
(A–D) Chromatograms (A and C) and quantitation of fold change (B and D) of various N-acyl Phes (A and B) or various C18:1-amino acids (C and D) in plasma of male C57BL/6 mice in thermoneutrality after tail vein injection of AAV-GFP or AAV-PM20D1 by targeted MRM. Mice were placed into 30°C at 6 weeks old, injected with virus at 7 weeks old, and high fat diet (HFD) was started 7 days post injection. Mice were maintained at 30°C for the duration of the experiment. The comparative targeted metabolomics was performed on plasma harvested 47 days post injection. For (A) and (C), chromatograms are from one representative mouse per group. For (B) and (D), the absolute quantitation in AAV-GFP versus AAV-PM20D1 is as follows: C16-Phe, 6 versus 15 nM, respectively; C18:2-Phe, 2 versus 7 nM, respectively; C18:1-Phe, 4 versus 10 nM, respectively; C18:1-Leu, 6 versus 29 nM, respectively.
(E) Fold change (cold/room temperature) of the indicated plasma N-acyl amino acids following cold exposure for the indicated times by targeted MRM.
For (B) and (D–E), n=4–5/group, mean ± SEM, * p<0.05, ** p<0.01.