A Fast and Accurate Method to Identify and Quantify Enzymes in Brush-Border Membranes: In Situ Hydrolysis Followed by Nano LC-MS/MS

Brun, Antonio; Magallanes, M; Rio, Carlos Martı́nez del; Barrett-Wilt, Gregory A.; Karasov, William H.; Caviedes‐Vidal, Enrique

doi:10.3390/mps3010015

Cited by 7 publications

(3 citation statements)

References 17 publications

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“…Sucrase activity is owing to the action of a membrane-bound alpha-glucosidase enzyme called sucrase-isomaltase (coded by the SI gene) and expressed in the apical membrane of intestinal cells (called the brush-border membrane, Brun et al., 2020a ). Recently developed methods in proteomics allow quantifying the abundance of different digestive enzymes in the brush-border membrane ( Brun et al., 2020b ). These proteomics methods have revealed that in the muscicapoid lineage (predominantly insectivorous) that includes starlings, thrushes, and mockingbirds, sucrase is present ( Brun et al., 2020a ) but has lost the ability to hydrolyze sucrose ( Martínez del Rio, 1990b ).…”

Section: Resultsmentioning

confidence: 99%

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents

et al. 2021

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Summary The major lineages of nectar-feeding birds (hummingbirds, sunbirds, honeyeaters, flowerpiercers, and lorikeets) are considered examples of convergent evolution. We compared sucrose digestion capacity and sucrase enzymatic activity per unit intestinal surface area among 50 avian species from the New World, Africa, and Australia, including 20 nectarivores. With some exceptions, nectarivores had smaller intestinal surfaces, higher sucrose hydrolysis capacity, and greater sucrase activity per unit intestinal area. Convergence analysis showed high values for sucrose hydrolysis and sucrase activity per unit intestinal surface area in specialist nectarivores, matching the high proportion of sucrose in the nectar of the plants they pollinate. Plants pollinated by generalist nectar-feeding birds in the Old and New Worlds secrete nectar in which glucose and fructose are the dominant sugars. Matching intestinal enzyme activity in birds and nectar composition in flowers appears to be an example of convergent coevolution between plants and pollinators on an intercontinental scale.

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Section: Resultsmentioning

confidence: 99%

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents

et al. 2021

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“…Several recent studies have used the state-of-the-art mass spectrometry approach to identify cancer-specific biomarkers in human diseases [ 84 , 85 ] and additional analyses of proteomics data led to the identification of the specific pathways involved in mediating carcinogenesis [ 86 ]. Wei Chu et al identified oral cancer specific biomarkers [ 87 ] using mass spectrometry in oral cancer patients’ saliva.…”

Section: Cancer Therapy (Radiation)-induced Immune Modulationmentioning

confidence: 99%

Dark Side of Cancer Therapy: Cancer Treatment-Induced Cardiopulmonary Inflammation, Fibrosis, and Immune Modulation

Boopathi

Thangavel

2021

IJMS

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Advancements in cancer therapy increased the cancer free survival rates and reduced the malignant related deaths. Therapeutic options for patients with thoracic cancers include surgical intervention and the application of chemotherapy with ionizing radiation. Despite these advances, cancer therapy-related cardiopulmonary dysfunction (CTRCPD) is one of the most undesirable side effects of cancer therapy and leads to limitations to cancer treatment. Chemoradiation therapy or immunotherapy promote acute and chronic cardiopulmonary damage by inducing reactive oxygen species, DNA damage, inflammation, fibrosis, deregulation of cellular immunity, cardiopulmonary failure, and non-malignant related deaths among cancer-free patients who received cancer therapy. CTRCPD is a complex entity with multiple factors involved in this pathogenesis. Although the mechanisms of cancer therapy-induced toxicities are multifactorial, damage to the cardiac and pulmonary tissue as well as subsequent fibrosis and organ failure seem to be the underlying events. The available biomarkers and treatment options are not sufficient and efficient to detect cancer therapy-induced early asymptomatic cell fate cardiopulmonary toxicity. Therefore, application of cutting-edge multi-omics technology, such us whole-exome sequencing, DNA methylation, whole-genome sequencing, metabolomics, protein mass spectrometry and single cell transcriptomics, and 10 X spatial genomics, are warranted to identify early and late toxicity, inflammation-induced carcinogenesis response biomarkers, and cancer relapse response biomarkers. In this review, we summarize the current state of knowledge on cancer therapy-induced cardiopulmonary complications and our current understanding of the pathological and molecular consequences of cancer therapy-induced cardiopulmonary fibrosis, inflammation, immune suppression, and tumor recurrence, and possible treatment options for cancer therapy-induced cardiopulmonary toxicity.

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“…Trypsin digested proteins were identified by LC-ESI-QTOF as per the standard process described elsewhere (Kaplan et al, 2004;Brun et al, 2020). Briefly, trypsin shaved proteins obtained from cell surface or ECM were treated with DTT and iodoacetamide for 1 h and the samples were digested with trypsin (1 µg/ml) for another 12-16 h at 37 • C. The trypsin enzyme was inactivated by adding 5% Tri-Fluro acetic acid (TFA) to the peptide mixtures in tube.…”

Section: Identification Of Proteins By Lc-esi-qtofmentioning

confidence: 99%

Glabridin Averts Biofilms Formation in Methicillin-Resistant Staphylococcus aureus by Modulation of the Surfaceome

2020

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Staphylococcus aureus is an opportunistic bacterium of the human body and a leading cause of nosocomial infections. Methicillin resistant S. aureus (MRSA) infections involving biofilm lead to higher mortality and morbidity in patients. Biofilm causes serious clinical issues, as it mitigates entry of antimicrobials to reach the etiological agents. It plays an important role in resilient chronic infections which place an unnecessary burden on antibiotics and the associated costs. To combat drug-resistant infection involving biofilm, there is a need to discover potential anti-biofilm agents. In this study, activity of polyphenolic flavonoid glabridin against biofilm formation of methicillin resistant clinical isolates of S. aureus is being reported for the first time. Crystal violet assay and scanning electron microscopy evidences shows that glabridin prevents formation of cells clusters and attachment of methicillin resistant clinical isolate (MRSA 4423) of S. aureus to the surface in a dose dependent manner. Gel free proteomic analysis of biofilm matrix by LC-ESI-QTOF confirmed the existence of several proteins known to be involved in cells adhesion. Furthermore, expression analysis of cell surface proteins revealed that glabridin significantly down regulates an abundance of several surface-associated adhesins including fibronectin binding proteins (FnbA, FnbB), serineaspartate repeat-containing protein D (SdrD), immunoglobulin-binding protein G (Sbi), and other virulence factors which were induced by extracellular glucose in MRSA 4423. In addition, several moonlighting proteins (proteins with multiple functions) such as translation elongation factors (EF-Tu, EF-G), chaperone protein (DnaK), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and pyruvate kinase (PK) were detected on the cell surface wherein their abundance was inversely proportional to surface-associated adhesins. This study clearly suggests that glabridin prevents biofilm formation in S. aureus through modulation of the cell surface proteins.

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A Fast and Accurate Method to Identify and Quantify Enzymes in Brush-Border Membranes: In Situ Hydrolysis Followed by Nano LC-MS/MS

Cited by 7 publications

References 17 publications

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents

Dark Side of Cancer Therapy: Cancer Treatment-Induced Cardiopulmonary Inflammation, Fibrosis, and Immune Modulation

Glabridin Averts Biofilms Formation in Methicillin-Resistant Staphylococcus aureus by Modulation of the Surfaceome

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