Effect of Protein Denaturation and Enzyme Inhibitors on Proteasomal-Mediated Production of Peptides in Human Embryonic Kidney Cells

Dasgupta, Sayani; Fishman, Michael; Castro, Leandro M.; Ferro, Emer S.; Fricker, Lloyd D.

doi:10.3390/biom9060207

Cited by 8 publications

(9 citation statements)

References 66 publications

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“…LC-MS and LC-MS/MS experiments were performed in an Orbitrap (OT) Fusion Tribrid mass spectrometer (Thermo Fisher Scientific, Waltham, MA, U.S.A) within the facilities at Harvard Medical School while using similar conditions to those previously described [81]. Briefly, OT top speed data-dependent precursor ion selection was used.…”

Section: Mass Spectrometry and Data Analysesmentioning

confidence: 99%

The Relevance of Thimet Oligopeptidase in the Regulation of Energy Metabolism and Diet-Induced Obesity

et al. 2020

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Thimet oligopeptidase (EC 3.4.24.15; EP24.15; THOP1) is a potential therapeutic target, as it plays key biological functions in processing biologically functional peptides. The structural conformation of THOP1 provides a unique restriction regarding substrate size, in that it only hydrolyzes peptides (optimally, those ranging from eight to 12 amino acids) and not proteins. The proteasome activity of hydrolyzing proteins releases a large number of intracellular peptides, providing THOP1 substrates within cells. The present study aimed to investigate the possible function of THOP1 in the development of diet-induced obesity (DIO) and insulin resistance by utilizing a murine model of hyperlipidic DIO with both C57BL6 wild-type (WT) and THOP1 null (THOP1−/−) mice. After 24 weeks of being fed a hyperlipidic diet (HD), THOP1−/− and WT mice ingested similar chow and calories; however, the THOP1−/− mice gained 75% less body weight and showed neither insulin resistance nor non-alcoholic fatty liver steatosis when compared to WT mice. THOP1−/− mice had increased adrenergic-stimulated adipose tissue lipolysis as well as a balanced level of expression of genes and microRNAs associated with energy metabolism, adipogenesis, or inflammation. Altogether, these differences converge to a healthy phenotype of THOP1−/− fed a HD. The molecular mechanism that links THOP1 to energy metabolism is suggested herein to involve intracellular peptides, of which the relative levels were identified to change in the adipose tissue of WT and THOP1−/− mice. Intracellular peptides were observed by molecular modeling to interact with both pre-miR-143 and pre-miR-222, suggesting a possible novel regulatory mechanism for gene expression. Therefore, we successfully demonstrated the previously anticipated relevance of THOP1 in energy metabolism regulation. It was suggested that intracellular peptides were responsible for mediating the phenotypic differences that are described herein by a yet unknown mechanism of action.

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Section: Mass Spectrometry and Data Analysesmentioning

confidence: 99%

The Relevance of Thimet Oligopeptidase in the Regulation of Energy Metabolism and Diet-Induced Obesity

et al. 2020

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“…The majority of HEK293 and and human neuroblastoma SH-SY5Y cell peptides identified during peptidome studies were likely to be generated by proteasomes, as they are fragments of intracellular proteins, mostly containing from 5 to 17 amino acids in length and having either a hydrophobic (75%), basic (12%), or acidic (8%) amino acids at their C terminus [ 95 ]. Thus, InPeps’ profile was shown to be strongly modulated by proteasome inhibition, both in HEK293 and SH-SY5Y cells and murine brain [ 72 , 80 , 81 , 96 , 97 , 98 , 99 , 100 ]. When considering the length of the endogenous substrates of THOP1, the majority of the substrates were peptides that were 9 to 11 amino acids long.…”

Section: Seminal Experiments Evidencing That Intracellular Peptidementioning

confidence: 99%

Thimet Oligopeptidase Biochemical and Biological Significances: Past, Present, and Future Directions

2020

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Thimet oligopeptidase (EC 3.4.24.15; EP24.15, THOP1) is a metallopeptidase ubiquitously distributed in mammalian tissues. Beyond its previously well characterized role in major histocompatibility class I (MHC-I) antigen presentation, the recent characterization of the THOP1 C57BL6/N null mice (THOP1−/−) phenotype suggests new key functions for THOP1 in hyperlipidic diet-induced obesity, insulin resistance and non-alcoholic liver steatosis. Distinctive levels of specific intracellular peptides (InPeps), genes and microRNAs were observed when comparing wild type C57BL6/N to THOP1−/− fed either standard or hyperlipidic diets. A possible novel mechanism of action was suggested for InPeps processed by THOP1, which could be modulating protein-protein interactions and microRNA processing, thus affecting the phenotype. Together, research into the biochemical and biomedical significance of THOP1 suggests that degradation by the proteasome is a step in the processing of various proteins, not merely for ending their existence. This allows many functional peptides to be generated by proteasomal degradation in order to, for example, control mRNA translation and the formation of protein complexes.

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“…These studies also demonstrated that the overexpression of THOP1, which has been shown to be involved in the metabolism of specific intracellular peptides [53] and modulation of signal transduction of angiotensin II (AT1) and β-adrenergic GPCR in CHO and HEK293 cells. Recent studies in HEK293 cells showed that most of these intracellular peptides are generated by the proteasome, where additional intracellular peptide-generating peptidases also exist [52,[54][55][56].…”

Section: Brief Historical Perspective On Proteasome-derived Intracellmentioning

confidence: 99%

“…Intracellular peptides are mainly generated by proteasome [55,56,[60][61][62][63] and should function inside the cells, possibly regulating protein interactions [43,54,64,65]. However, a significant portion of intracellular peptides can be secreted [66], suggesting that they can also act on membrane receptors [67,68].…”

Section: Brief Historical Perspective On Proteasome-derived Intracellmentioning

confidence: 99%

Peptides from Natural or Rationally Designed Sources Can Be Used in Overweight, Obesity, and Type 2 Diabetes Therapies

et al. 2020

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Overweight and obesity are among the most prominent health problems in the modern world, mostly because they are either associated with or increase the risk of other diseases such as type 2 diabetes, hypertension, and/or cancer. Most professional organizations define overweight and obesity according to individual body–mass index (BMI, weight in kilograms divided by height squared in meters). Overweight is defined as individuals with BMI from 25 to 29, and obesity as individuals with BMI ≥30. Obesity is the result of genetic, behavioral, environmental, physiological, social, and cultural factors that result in energy imbalance and promote excessive fat deposition. Despite all the knowledge concerning the pathophysiology of obesity, which is considered a disease, none of the existing treatments alone or in combination can normalize blood glucose concentration and prevent debilitating complications from obesity. This review discusses some new perspectives for overweight and obesity treatments, including the use of the new orally active cannabinoid peptide Pep19, the advantage of which is the absence of undesired central nervous system effects usually experienced with other cannabinoids.

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Effect of Protein Denaturation and Enzyme Inhibitors on Proteasomal-Mediated Production of Peptides in Human Embryonic Kidney Cells

Cited by 8 publications

References 66 publications

The Relevance of Thimet Oligopeptidase in the Regulation of Energy Metabolism and Diet-Induced Obesity

The Relevance of Thimet Oligopeptidase in the Regulation of Energy Metabolism and Diet-Induced Obesity

Thimet Oligopeptidase Biochemical and Biological Significances: Past, Present, and Future Directions

Peptides from Natural or Rationally Designed Sources Can Be Used in Overweight, Obesity, and Type 2 Diabetes Therapies

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