Fig. 1.

PLIN5 is a substrate for in vitro phosphorylation by PKA. A: Amino acid sequence of a portion of PLIN5 are aligned for homo sapiens (135–151), pan (148–164), rattus (150–166), mus (150–166), chiroptera (182–198), and ovis aries (212–228). The sequences show conservation at the PKA consensus sequence corresponding to murine S155 and putative sites at S161 and S163. B: Purified recombinant murine PLIN5 was incubated with PKA in vitro, the peptides were digested and phospho-peptides detected by LC-MS/MS. Data were reproduced in three independent experiments. C: Recombinant murine PLIN5 was incubated with the catalytic subunit of PKA and [γ-³²P]ATP followed by SDS-PAGE, transfer to a PVDF membrane and autoradiography. Representative autoradiograph (above) and PLIN5 immunoblot (below). D: Quantification of [γ-³²P]ATP incorporation into PLIN5; n = 2 per group. For panel (D), one-way ANOVA with Bonferroni post hoc analysis was performed. ∗P < 0.05 versus PLIN5 + PKA.