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GNPS - Feline fecal samples (quantification of polyamine conjugated to bile acids)

ABSTRACT: Fecal samples from felines extracted with 50:50 MeOH:H2O and chromatographed using a Phenomenex polar C18 column. MS/MS data was acquired on a QExactive Orbitrap in positive ionization mode. Quantification of polyamines conjugated to bile acids was performed (cholyl-putrescine, cholyl-cadaverine, cholyl-N-acetyl-putrescine), in addition to taurocholic acid and glycocholic acid.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Panthera Tigris (ncbitaxon:9694) Leptailurus Serval (ncbitaxon:61405) Panthera Onca (ncbitaxon:9690) Acinonyx Jubatus (ncbitaxon:32536) Herpailurus Yagouaroundi Panthera Leo (ncbitaxon:9689)

SUBMITTER: Pieter C Dorrestein

PROVIDER: MSV000093618 | GNPS | Sat Dec 09 06:46:00 GMT 2023

REPOSITORIES: GNPS

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Project description:Polyamines are aliphatic polycations that have emerged as important determinants of cell growth and viability in rapidly proliferating cells, including in the pathogenic protozoan parasite Leishmania donovani. In L. donovani, the polyamine spermidine is synthesized by the successive conversion of ornithine into putrescine (catalyzed by ornithine decarboxylase or ODC) and putrescine into spermidine (catalyzed by spermidine synthase or SPDSYN). Deletion of either ODC (del-odc) or SPDSYN (del-spdsyn) from the L. donovani genome renders these parasites auxotrophic for polyamines and these mutants are impaired in their ability to survive both in culture and within the mammalian host without the addition of exogenous polyamine supplementation. Significantly, del-odc parasites immediately cease proliferation after putrescine is removed from the culture media and perish within two weeks, while spermidine starved del-spdsyn mutants, which retain intracellular putrescine pools, show a slow-growth phenotype, and persist for several weeks in culture. To elucidate the key differences within the proteome of putrescine-starved del-odc cells and spermidine-starved del-spdsyn parasites, a shotgun quantitative proteomics approach was undertaken using TMT labeling and LC-MS/MS analysis. Briefly, three biological replicates each for mid-log phase del-odc and del-spdsyn promastigotes grown in the presence of exogenous putrescine (for del-odc) or spermidine (for del-spdsyn) supplementation were washed to remove the exogenous polyamine supplementation and incubated in polyamine-free media. At 24 and 48 h, cells from each biological replicate were isolated and prepared for tandem mass tag (TMT) labeling and downstream LC-MS/MS analyses. Peptides were identified using a database generated from the reference genome of L. donovani BPK282A1 strain. Changes in relative protein abundance for the polyamine-starved del-odc and del-spdsyn cell lines at 24 and 48 h were calculated by comparing aggregate total reporter ion intensities for each protein to that of the corresponding polyamine-supplemented 0-h timepoint.

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GNPS - Feline fecal samples (quantification of polyamine conjugated to bile acids)

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