Journal: Cell Reports

Article Title: The hepatic compensatory response to elevated systemic sulfide promotes diabetes

doi: 10.1016/j.celrep.2021.109958

Figure Lengend Snippet: Tst deletion results in increased hepatic sulfur excretion and a reduction of protein persulfidation (A) Schematic showing mammalian metabolism of hydrogen sulfide. The canonical production enzymes are shown in the cytosol. MPST, mercaptopyruvate sulfurtransferase; CBS, cystathionine beta synthase; CTH, cystathionine gamma lyase. Mitochondrial oxidation and disposal of hydrogen sulfide occurs through the SOP through the actions of SQOR (sulfide quinone oxidoreductase), ETHE1 (PDO), TST (thiosulfate sulfurtransferase), and SUOX (sulfite oxidase). These seven enzymes are major contributors to intracellular sulfide (and other inorganic sulfur) metabolism. For simplicity, the diagram does not include sulfide production, which can occur within mitochondria, or disposal pathways in the cytosol. The identity of oxidized sulfur species produced by SQOR remain disputed. The precise role of TST and other enzymes shown here remains under investigation. (B) Cysteine concentrations (MBB-HPLC) in medium incubated with primary hepatocytes in the presence (hatched pattern) or absence (no pattern) of 1 mM methionine from C57BL/6J (white bars, n = 4/treatment) and Tst −/− (red bars, n = 4/treatment) mice. (C) GSH concentrations (MBB-HPLC) in medium incubated with primary hepatocytes in the presence (hatched pattern) or absence (no pattern) of 1 mM methionine from C57BL/6J (white bars, n = 4/treatment) and Tst −/− (red bars, n = 4/treatment) mice. (D) Pie chart depicting the proportion of liver peptides that are significantly higher (82 peptides, purple space) or lower (311 peptides, yellow space) in their persulfidation rate in Tst −/− (n = 3) relative to C57BL/6J (n = 3) mice. (E) Total DTT-released cysteine-persulfidated liver protein as measured by REVERT total protein stain following western blotting, normalized to the total input protein of the sample from Tst −/− (red bar, n = 4) and C57BL/6J (white bar, n = 4) mice. Data with error bars are represented as mean ± SEM. Significance was calculated using 2-way ANOVA (B and C) or Student’s t test (E); ∗ p < 0.05, ∗∗ p < 0.01. For (B) and (C), the 2-way ANOVA reveals a main effect of genotype, indicated by ∗ or ∗∗ on the histogram. A significant effect of methionine was also found for (B) and (C), not indicated on the histogram. For (D), peptides were selected as being significant at a P-diff of 0.95 or greater. See also Figure S3 and .

Article Snippet: Primary antibodies used were; TST, Rabbit, GeneTex, GTX114858, MPST, Rabbit, Abcam, Ab224043, GOT1, Rabbit, Abcam, ab170950, GSTT1, Rabbit, Proteintech, 15838-1-AP, MAT1A, Rabbit, Abcam, ab129176, BHMT Rabbit, Proteintech, 15965-1-AP, CSAD, Rabbit, Abcam, ab91016, PPCS Rabbit, Atlas Antibodies, HPA031361.

Techniques: Produced, Incubation, Staining, Western Blot

Journal: Cell Reports

Article Title: The hepatic compensatory response to elevated systemic sulfide promotes diabetes

doi: 10.1016/j.celrep.2021.109958

Figure Lengend Snippet:

Article Snippet: Primary antibodies used were; TST, Rabbit, GeneTex, GTX114858, MPST, Rabbit, Abcam, Ab224043, GOT1, Rabbit, Abcam, ab170950, GSTT1, Rabbit, Proteintech, 15838-1-AP, MAT1A, Rabbit, Abcam, ab129176, BHMT Rabbit, Proteintech, 15965-1-AP, CSAD, Rabbit, Abcam, ab91016, PPCS Rabbit, Atlas Antibodies, HPA031361.

Techniques: Recombinant, Protease Inhibitor, Marker, Staining, Sequencing, Modification, Electron Microscopy, Cholesterol Assay, Multiplex sample analysis, Knock-Out, Gene Expression, Software