Comparative LC-MSMS Analysis of Extracellular Vesicles Extracted from Mouse Caecum in Fasted vs. Fed Mice

Bacterial extracellular vesicles (BEVs) produced by members of the intestinal microbiota can contribute not only to digestion but also mediate microbe-host cell communication via the transfer of functional biomolecules to mammalian host cells. An unresolved question is what host factors and conditions influence BEV cargoes and how do they impact on host cell function? To address this question, we analysed and compared the proteome of BEVs released by the major human gastrointestinal tract (GIT) symbiont Bacteroides thetaiotaomicron (Bt) in vivo in fed versus fasted animals using nano-liquid chromatography with tandem mass spectrometry (LC-MSMS). Among the proteins whose abundance was negatively affected by fasting, nine of ten proteins of the serine protease family, including the regulatory protein dipeptidyl peptidase-4 (DPP-4), were significantly decreased in BEVs produced in the GIT of fasted animals. Strikingly, in ex-tracellular vesicles produced by the intestinal epithelium of the same fasted mice, the proteins with the most increased abundance were serine protease inhibitors (serpins). Together, these findings suggest a dynamic interaction between GI bacteria and the host. Additionally, they indicate a regulatory role for the host in determining the balance be-tween bacterial serine proteases and host serpins exported in bacterial and host extra-cellular vesicles.