Quantitative Proteomics Using 15N SILAC Mouse

Koller, Antonius; Wen, Ruofeng; Wu, Xiao; Relucio, Jenne; Colognato, Holly; Chen, Emily I.

doi:10.14302/issn.2326-0793.jpgr-13-252

Cited by 4 publications

(3 citation statements)

References 30 publications

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“…RNAs that failed to have at least ten samples with CPM values above −3 were removed. b Linear plots representing log2(CPM) values with age for the top ten RNAs ranked by the lowest p -value (two-sided Wald test, unadjusted) 97 – 108 . …”

Section: Resultsmentioning

confidence: 99%

Skeletal muscle transcriptome in healthy aging

et al. 2021

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Age-associated changes in gene expression in skeletal muscle of healthy individuals reflect accumulation of damage and compensatory adaptations to preserve tissue integrity. To characterize these changes, RNA was extracted and sequenced from muscle biopsies collected from 53 healthy individuals (22–83 years old) of the GESTALT study of the National Institute on Aging–NIH. Expression levels of 57,205 protein-coding and non-coding RNAs were studied as a function of aging by linear and negative binomial regression models. From both models, 1134 RNAs changed significantly with age. The most differentially abundant mRNAs encoded proteins implicated in several age-related processes, including cellular senescence, insulin signaling, and myogenesis. Specific mRNA isoforms that changed significantly with age in skeletal muscle were enriched for proteins involved in oxidative phosphorylation and adipogenesis. Our study establishes a detailed framework of the global transcriptome and mRNA isoforms that govern muscle damage and homeostasis with age.

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Section: Resultsmentioning

confidence: 99%

Skeletal muscle transcriptome in healthy aging

et al. 2021

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“…2A ). The relative protein levels in the two cohorts were quantified by two-dimensional LC-MS/MS with stable-isotope amino acids labeled reference tissue lysate as described ( Koller et al , 2013 ; Sinnamon et al , 2012 ). We identified 1,346 proteins from WT and Mmp14 KO anlagen.…”

Section: Resultsmentioning

confidence: 99%

New insight into the role of MMP14 in metabolic balance

Mori

Bhat

Bruni‐Cardoso

et al. 2016

PeerJ

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Membrane-anchored matrix metalloproteinase 14 (MMP14) is involved broadly in organ development through both its proteolytic and signal-transducing functions. Knockout of Mmp14 (KO) in mice results in a dramatic reduction of body size and wasting followed by premature death, the mechanism of which is poorly understood. Since the mammary gland develops after birth and is thus dependent for its functional progression on systemic and local cues, we chose it as an organ model for understanding why KO mice fail to thrive. A global analysis of the mammary glands’ proteome in the wild type (WT) and KO mice provided insight into an unexpected role of MMP14 in maintaining metabolism and homeostasis. We performed mass spectrometry and quantitative proteomics to determine the protein signatures of mammary glands from 7 to 11 days old WT and KO mice and found that KO rudiments had a significantly higher level of rate-limiting enzymes involved in catabolic pathways. Glycogen and lipid levels in KO rudiments were reduced, and the circulating levels of triglycerides and glucose were lower. Analysis of the ultrastructure of mammary glands imaged by electron microscopy revealed a significant increase in autophagy signatures in KO mice. Finally, Mmp14 silenced mammary epithelial cells displayed enhanced autophagy. Applied to a systemic level, these findings indicate that MMP14 is a crucial regulator of tissue homeostasis. If operative on a systemic level, these findings could explain how Mmp14KO litter fail to thrive due to disorder in metabolism.

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“…2a). The relative protein levels in the two cohorts were quantified by two-dimensional LC-MS/MS with stableisotope amino acids labeled reference tissue lysate as described (Koller et al 2013;Sinnamon et al 2012). We identified 1346 proteins from WT and Mmp14KO anlagen.…”

Section: Proteomic Profile Of Mammary Glands Of Mmp14ko Reveals Defecmentioning

confidence: 99%

Peer Review #2 of "New insight into the role of MMP14 in metabolic balance (v0.1)"

2016

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Membrane-anchored matrix metalloproteinase 14 (MMP14) is involved broadly in organ development through both its proteolytic and signal-transducing functions. Knockout of Mmp14 (KO) in mice results in a dramatic reduction of body size and wasting followed by premature death, the mechanism of which is poorly understood. Since the mammary gland develops after birth and is thus dependent for its functional progression on systemic and local cues, we chose it as an organ model for understanding why KO mice fail to thrive. A global analysis of the mammary glands' proteome in the wild type (WT) and KO mice provided insight into an unexpected role of MMP14 in maintaining metabolism and homeostasis. We performed mass spectrometry and quantitative proteomics to determine the protein signatures of mammary glands from 7 to 11 days old WT and KO mice and found that KO rudiments had a significantly higher level of rate-limiting enzymes involved in catabolic pathways. Glycogen and lipid levels in KO rudiments were reduced, and the circulating levels of triglycerides and glucose were lower. Analysis of the ultrastructure of mammary glands imaged by electron microscopy revealed a significant increase in autophagy signatures in KO mice. Finally, Mmp14 silenced mammary epithelial cells displayed enhanced autophagy. Applied to a systemic level, these findings indicate that MMP14 is a crucial regulator of tissue homeostasis. If operative on a systemic level, these findings could explain how Mmp14 KO litter fail to thrive due to disorder inmetabolism.

show abstract

Quantitative Proteomics Using 15N SILAC Mouse

Cited by 4 publications

References 30 publications

Skeletal muscle transcriptome in healthy aging

Skeletal muscle transcriptome in healthy aging

New insight into the role of MMP14 in metabolic balance

Peer Review #2 of "New insight into the role of MMP14 in metabolic balance (v0.1)"

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