YoungJun Ju1, Ashley Untereiner2, Lingyun Wu2, Guangdong Yang3. 1. Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON P7B 5E1, Canada. 2. Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON P7B 5E1, Canada; Department of Health Science, Lakehead University, Thunder Bay, ON P7B 5E1, Canada. 3. Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON P7B 5E1, Canada; Department of Chemistry and Biochemistry, Laurentian University, Sudbury, ON P3E 2C6, Canada. Electronic address: gyang2@laurentian.ca.
Abstract
BACKGROUND: Cystathionine gamma-lyase (CSE)-derived hydrogen sulfide (H(2)S) possesses diverse roles in the liver, affecting lipoprotein synthesis, insulin sensitivity, and mitochondrial biogenesis. H(2)S S-sulfhydration is now proposed as a major mechanism for H(2)S-mediated signaling. Pyruvate carboxylase (PC) is an important enzyme for gluconeogenesis. S-sulfhydration regulation of PC by H(2)S and its implication in gluconeogenesis in the liver have been unknown. METHODS: Gene expressions were analyzed by real-time PCR and western blotting, and protein S-sulfhydration was assessed by both modified biotin switch assay and tag switch assay. Glucose production and PC activity was measured with coupled enzyme assays, respectively. RESULTS: Exogenously applied H(2)S stimulates PC activity and gluconeogenesis in both HepG2 cells and mouse primary liver cells. CSE overexpression enhanced but CSE knockout reduced PC activity and gluconeogenesis in liver cells, and blockage of PC activity abolished H(2)S-induced gluconeogenesis. H(2)S had no effect on the expressions of PC mRNA and protein, while H(2)S S-sulfhydrated PC in a dithiothreitol-sensitive way. PC S-sulfhydration was significantly strengthened by CSE overexpression but attenuated by CSE knockout, suggesting that H(2)S enhances glucose production through S-sulfhydrating PC. Mutation of cysteine 265 in human PC diminished H(2)S-induced PC S-sulfhydration and activity. In addition, high-fat diet feeding of mice decreased both CSE expression and PC S-sulfhydration in the liver, while glucose deprivation of HepG2 cells stimulated CSE expression. CONCLUSIONS: CSE/H(2)S pathway plays an important role in the regulation of glucose production through S-sulfhydrating PC in the liver. GENERAL SIGNIFICANCE: Tissue-specific regulation of CSE/H(2)S pathway might be a promising therapeutic target of diabetes and other metabolic syndromes.
BACKGROUND:Cystathionine gamma-lyase (CSE)-derived hydrogen sulfide (H(2)S) possesses diverse roles in the liver, affecting lipoprotein synthesis, insulin sensitivity, and mitochondrial biogenesis. H(2)S S-sulfhydration is now proposed as a major mechanism for H(2)S-mediated signaling. Pyruvate carboxylase (PC) is an important enzyme for gluconeogenesis. S-sulfhydration regulation of PC by H(2)S and its implication in gluconeogenesis in the liver have been unknown. METHODS: Gene expressions were analyzed by real-time PCR and western blotting, and protein S-sulfhydration was assessed by both modified biotin switch assay and tag switch assay. Glucose production and PC activity was measured with coupled enzyme assays, respectively. RESULTS: Exogenously applied H(2)S stimulates PC activity and gluconeogenesis in both HepG2 cells and mouse primary liver cells. CSE overexpression enhanced but CSE knockout reduced PC activity and gluconeogenesis in liver cells, and blockage of PC activity abolished H(2)S-induced gluconeogenesis. H(2)S had no effect on the expressions of PC mRNA and protein, while H(2)S S-sulfhydrated PC in a dithiothreitol-sensitive way. PC S-sulfhydration was significantly strengthened by CSE overexpression but attenuated by CSE knockout, suggesting that H(2)S enhances glucose production through S-sulfhydrating PC. Mutation of cysteine 265 in humanPC diminished H(2)S-induced PC S-sulfhydration and activity. In addition, high-fat diet feeding of mice decreased both CSE expression and PC S-sulfhydration in the liver, while glucose deprivation of HepG2 cells stimulated CSE expression. CONCLUSIONS:CSE/H(2)S pathway plays an important role in the regulation of glucose production through S-sulfhydrating PC in the liver. GENERAL SIGNIFICANCE: Tissue-specific regulation of CSE/H(2)S pathway might be a promising therapeutic target of diabetes and other metabolic syndromes.
Authors: Pauline M Snijder; Madina Baratashvili; Nicola A Grzeschik; Henri G D Leuvenink; Lucas Kuijpers; Sippie Huitema; Onno Schaap; Ben N G Giepmans; Jeroen Kuipers; Jan Lj Miljkovic; Aleksandra Mitrovic; Eelke M Bos; Csaba Szabó; Harm H Kampinga; Pascale F Dijkers; Eelke M Bos; Csaba Szabó; Harm H Kampinga; Pascale F Dijkers; Wilfred F A den Dunnen; Milos R Filipovic; Harry van Goor; Ody C M Sibon Journal: Mol Med Date: 2015-10-13 Impact factor: 6.354
Authors: Hui Peng; Yixiang Zhang; Lauren D Palmer; Thomas E Kehl-Fie; Eric P Skaar; Jonathan C Trinidad; David P Giedroc Journal: ACS Infect Dis Date: 2017-09-06 Impact factor: 5.084