Detecting Protein Carbonylation in Adipose Tissue and in Cultured Adipocytes

Xu, Qinghui; Hahn, Wendy S.; Bernlohr, David A.

doi:10.1016/b978-0-12-800280-3.00014-1

Cited by 6 publications

(3 citation statements)

References 24 publications

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“…Carbonylated total protein and smooth muscle actin-alpha measurement EZ-link hydrazine-biotin was used to label protein carbonyl as previously described (Grimsrud et al, 2007;Xu et al, 2014b). Small intestines were collected in ice-cold PBS following removal of intestinal contents.…”

Section: Tissue Carbonyl Assay Detectionmentioning

confidence: 99%

The asymmetric Pitx2 gene regulates gut muscular-lacteal development and protects against fatty liver disease

Mahadevan

Elysee

et al. 2021

Cell Reports

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SUMMARY Intestinal lacteals are essential lymphatic channels for absorption and transport of dietary lipids and drive the pathogenesis of debilitating metabolic diseases. However, organ-specific mechanisms linking lymphatic dysfunction to disease etiology remain largely unknown. In this study, we uncover an intestinal lymphatic program that is linked to the left-right (LR) asymmetric transcription factor Pitx2 . We show that deletion of the asymmetric Pitx2 enhancer ASE alters normal lacteal development through the lacteal-associated contractile smooth muscle lineage. ASE deletion leads to abnormal muscle morphogenesis induced by oxidative stress, resulting in impaired lacteal extension and defective lymphatic system-dependent lipid transport. Surprisingly, activation of lymphatic system-independent trafficking directs dietary lipids from the gut directly to the liver, causing diet-induced fatty liver disease. Our study reveals the molecular mechanism linking gut lymphatic function to the earliest symmetry-breaking Pitx2 and highlights the important relationship between intestinal lymphangiogenesis and the gut-liver axis.

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Section: Tissue Carbonyl Assay Detectionmentioning

confidence: 99%

The asymmetric Pitx2 gene regulates gut muscular-lacteal development and protects against fatty liver disease

Mahadevan

Elysee

et al. 2021

Cell Reports

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“…EZ-link hydrazine-biotin was used to label protein carbonyl as previously described [115, 116]. Small intestines were collected in ice-cold PBS following removal of intestinal contents.…”

Section: Methods Detailsmentioning

confidence: 99%

The asymmetric Pitx2 regulates intestinal muscular-lacteal development and protects against fatty liver disease

Kurpios

Mahadevan

et al. 2021

Preprint

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Intestinal lacteals are the essential lymphatic channels for absorption and transport of dietary lipids and drive pathogenesis of debilitating metabolic diseases. Yet, organ-specific mechanisms linking lymphatic dysfunction to disease etiology remain largely unknown. In this study, we uncover a novel intestinal lymphatic program that is linked to the left-right (LR) asymmetric transcription factor Pitx2. We show that deletion of the asymmetric Pitx2 enhancer, ASE, alters normal lacteal development through the lacteal-associated contractile smooth muscle lineage. ASE deletion leads to abnormal muscle morphogenesis induced by oxidative stress, resulting in impaired lacteal extension and defective lymphatic-dependent lipid transport. Surprisingly, activation of lymphatic-independent trafficking directs dietary lipids from the gut directly to the liver, causing diet-induced fatty liver disease. In summary, our studies reveal the molecular mechanism linking gut lymphatic development to the earliest symmetry-breaking Pitx2 and highlight the important relationship between intestinal lymphangiogenesis and gut-liver axis.

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“…This is not unusual for chemically driven modifications where enzymatic regulation is not a driving factor. Previous studies defining cytoplasmic or mitochondrial protein targets of carbonylation have been limited in depth due to the use of nonspecific derivatization methods that do not enable one to distinguish between sites of modification or the many possible types of carbonylation events (13,47). Our proteomic workflow ( Fig.…”

Section: Nuclear Carbonylation Of Zinc Finger Proteinsmentioning

confidence: 99%

Obesity-induced protein carbonylation in murine adipose tissue regulates the DNA-binding domain of nuclear zinc finger proteins

Hauck

Zhou

Hahn

et al. 2018

Journal of Biological Chemistry

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In obesity-linked insulin resistance, oxidative stress in adipocytes leads to lipid peroxidation and subsequent carbonylation of proteins by diffusible lipid electrophiles. Reduction in oxidative stress attenuates protein carbonylation and insulin resistance, suggesting that lipid modification of proteins may play a role in metabolic disease, but the mechanisms remain incompletely understood. Herein, we show that , diet-induced obesity in mice surprisingly results in preferential carbonylation of nuclear proteins by 4-hydroxy--2,3-nonenal (4-HNE) or 4-hydroxy--2,3-hexenal (4-HHE). Proteomic and structural analyses revealed that residues in or around the sites of zinc coordination of zinc finger proteins, such as those containing the C2H2 or MATRIN, RING, C3H1, or N4-type DNA-binding domains, are particularly susceptible to carbonylation by lipid aldehydes. These observations strongly suggest that carbonylation functionally disrupts protein secondary structure supported by metal coordination. Analysis of one such target, the nuclear protein estrogen-related receptor γ (ERR-γ), showed that ERR-γ is modified by 4-HHE in the obese state. carbonylation decreased the DNA-binding capacity of ERR-γ and correlated with the obesity-linked down-regulation of many key genes promoting mitochondrial bioenergetics. Taken together, these findings reveal a novel mechanistic connection between oxidative stress and metabolic dysfunction arising from carbonylation of nuclear zinc finger proteins, such as the transcriptional regulator ERR-γ.

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Detecting Protein Carbonylation in Adipose Tissue and in Cultured Adipocytes

Cited by 6 publications

References 24 publications

The asymmetric Pitx2 gene regulates gut muscular-lacteal development and protects against fatty liver disease

The asymmetric Pitx2 gene regulates gut muscular-lacteal development and protects against fatty liver disease

The asymmetric Pitx2 regulates intestinal muscular-lacteal development and protects against fatty liver disease

Obesity-induced protein carbonylation in murine adipose tissue regulates the DNA-binding domain of nuclear zinc finger proteins

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