297 research outputs found
Proteomics of Hazelnut (Corylus avellana)
No full textHazelnut (Corylus avellana) is widely used in food production in raw, roasted, salted, and paste form. Proteins are key components conferring favorable sensory, technological, and functional properties to hazelnut. For these reasons, the protein fraction of hazelnut has been the subject of extensive research over the last years. In addition, the complete annotation of the hazelnut proteome at the molecular level is of great interest, in particular in clinical research because hazelnut proteins can elicit even severe allergic reactions in sensitive subjects. This chapter provides an overview of the current knowledge on the hazelnut proteome
Proteomic Analysis of Beer
No full textProteins determine several key sensory and technological properties of beer. Most of the protein components of beer are barley pathogenesis–related proteins, which endure the harsh brewing treatments. Due to their intrinsic resistance, they can also survive gastrointestinal digestion in humans, evoking immune responses in predisposed individuals. For these reasons, beer proteins have been studied for several decades. However, only the recent development of the mass spectrometry–based proteomic techniques is disclosing the “deep” proteome of beer. Several dozens of barley- and yeast-derived proteins constitute the current “core” proteome of beer, while many others are specific to beer brands or styles. Proteolytic peptides of beer include a multitude of sequences derived from prolamins that are potentially harmful for celiacs. In this chapter, we survey the compositional and functional aspects of the beer proteome/peptidome emphasizing the most recent findings obtained using the newest proteomic approaches
A mass spectrometric assay for the quantification of neuropeptide PYY in plasma
No full textA multiple reaction monitoring mass spectrometric assay for the quantification of PYY in human plasma has been developed. A two stage sample preparation protocol was employed in which plasma containing the full length neuropeptide was first digested using trypsin, followed by solid-phase extraction to extract the digested peptide from the complex plasma matrix. The peptide extracts were analysed by LC-MS using multiple reaction monitoring to detect and quantify PYY. The method has been validated for plasma samples, yielding linear responses over the range 5–1,000 ng mL−1. The method is rapid, robust and specific for plasma PYY detection
Integrative Proteomic Analysis of Digestive Tract Glycosidases from the Invasive Golden Apple Snail, Pomacea canaliculata
No full textThe freshwater snail Pomacea canaliculata, an invasive species of global significance, possesses a well-developed digestive system and diverse feeding mechanisms enabling the intake of a wide variety of food. The identification of glycosidases in adult snails would increase the understanding of their digestive physiology and potentially generate new opportunities to eradicate and/or control this invasive species. In this study, liquid chromatography coupled to tandem mass spectrometry was applied to define the occurrence, diversity, and origin of glycoside hydrolases along the digestive tract of P. canaliculata. A range of cellulases, hemicellulases, amylases, maltases, fucosidases, and galactosidases were identified across the digestive tract. The digestive gland and the contents of the crop and style sac yield a higher diversity of glycosidase-derived peptides. Subsequently, peptides derived from 81 glycosidases (46 proteins from the public database and 35 uniquely from the transcriptome database) that were distributed among 13 glycoside hydrolase families were selected and quantified using multiple reaction monitoring mass spectrometry. This study showed a high glycosidase abundance and diversity in the gut contents of P. canaliculata which participate in extracellular digestion of complex dietary carbohydrates. Salivary and digestive glands were the main tissues involved in their synthesis and secretion.Fil: Escobar Correas, Sophia Melanie. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Fisiología; ArgentinaFil: Mendoza Porras, Omar. Commonwealth Scientific And Industrial Research Organization; AustraliaFil: Dellagnola, Federico A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Colgrave, Michelle L.. Commonwealth Scientific And Industrial Research Organization; AustraliaFil: Vega, Israel Aníbal. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Universidad Nacional de Cuyo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentin
Challenges in mass spectrometry-based quantification of bioactive peptides: A case study exploring the neuropeptide Y family
No full textThe study of biologically active peptides is critical to the understanding of physiological pathways, especially those involved in the development of disease. Historically, the measurement of biologically active endogenous peptides has been undertaken by radioimmunoassay, a highly sensitive and robust technique that permits the detection of physiological concentrations in different biofluid and tissue extracts. Over recent years, a range of mass spectrometric approaches have been applied to peptide quantification with limited degrees of success. Neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) belong to the NPY family exhibiting regulatory effects on appetite and feeding behavior. The physiological significance of these peptides depends on their molecular forms and in vivo concentrations systemically and at local sites within tissues. In this report, we describe an approach for quantification of individual peptides within mixtures using high-performance liquid chromatography electrospray ionization tandem mass spectrometry analysis of the NPY family peptides. Aspects of quantification including sample preparation, the use of matrix-matched calibration curves, and internal standards will be discussed. This method for the simultaneous determination of NPY, PYY, and PP was accurate and reproducible but lacks the sensitivity required for measurement of their endogenous concentration in plasma. The advantages of mass spectrometric quantification will be discussed alongside the current obstacles and challenges. © 2012 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 98: 357–366, 2012.\u
In vitro transport and satiety of a beta-lactoglobulin dipeptide and beta-casomorphin-7 and its metabolites
No full textFirst published online 19 May 2014Understanding the digestive behaviour and biological activities of dairy proteins may help to develop model dairy products with targeted health outcomes including increased satiety and healthy weight maintenance. Caseins and whey proteins constitute over 95% of milk proteins with consumption of these proteins associated with increased satiety and a decreased prevalence of metabolic disorders. To investigate the in vitro digestive behaviour and satiety of dairy proteins at the intestinal epithelium, the in vitro transport and hydrolysis of 500-2000 μM β-casomorphin-7 (YPFPGPI or β-CM7) and a β-lactoglobulin (β-Lg) dipeptide (YL) was measured using Caco-2 cell monolayers grown on transwells as a model of the intestinal epithelium. Transport of YL was concentration dependent and ranged from 0.37-5.26 × 10(-6) cm s(-1), whereas transport of β-CM7 was only detected at 2000 μM and was significantly lower at 0.13 × 10(-6) cm s(-1). Rapid hydrolysis of β-CM7 in the apical chamber by the Caco-2 cells produced three peptide metabolites: YP, GPI and FPGPI. All of these metabolites were detected in the basolateral chamber after 30 min with both the YP and GPI peptides transporting at a higher rate than intact β-CM7. In vitro satiety was indicated by the secretion of cholecystokinin [26-33] (CCK-8) and glucagon-like peptide 1 (GLP-17-36NH2) in the STC-1 enteroendocrine cell model. CCK-8 secretion was highest in response to β-CM7 followed by the β-CM7 metabolite FPGPI. CCK-8 secretion however was not significantly stimulated by the tri- or dipeptides. Secretion of GLP-1 was not significantly stimulated by β-CM7 or YL. These in vitro results suggest that dairy peptide size enhances CCK-8 secretion, whilst limiting transport across Caco-2 monolayers.Simone Osborne, Wei Chen, Rama Addepalli, Michelle Colgrave, Tanoj Singh, Cuong Tran and Li Da
Optimisation of the extraction methods for proteomics analysis of three narrow-leafed lupin cultivars
No full textIn this study the proteome composition of three narrow-leafed lupin varieties (Tanjil, Unicrop, and P27255) resulted form four extraction methods was studied using discovery proteomics, this was followed by targeted quantitative analysis of the major lupin seed storage proteins known as conglutins.\nThe collection includes the following data:\n1- The information-dependent acquisition mass spectrometry (IDA-MS) data acquired on acquired on TripleTOF 6600 (SCIEX) mass\n2- ProteinPilot Reports for the IDA data which includes information about the data quality and protein and peptide identifications\n3-multiple reaction monitoring-mass spectrometry (LC-MRM-MS) data acquired on a 6500 QTRAP mass spectrometer (SCIEX)\n\nLineage: Proteome data were acquired using a liquid chromatograph-mass spectrometer (Eksigent nanoLC 415 - SCIEX 6600 QqTOF). Samples were separated using a C18 chromatography column and the eluting peptides measured by the mass spectrometer as mass-to-charge ratios. The mass spectrometer was operated in information-dependant acquisition mode whereby the machine surveys precursor ions and then selects mass-to-charge ratios for fragmentation in repeating cycles until the liquid chromatography program has completed
Mass spectrometry study of the white lupin allergens in raw and fermented samples
No full textThe changes in the protein profiles of white lupin as result of fermentation induced by Rhizopus oligosporus was evaluated using discovery proteomic analysis. Moreover, the changes in the abundances of allergen derived tryptic peptides were evaluated using targeted quantitative proteomics study. This collection includes the following data files:\nThe IDA-MS data, ProteinPilot Reports for the IDA data database search, and MRM data.\n\nLineage: Discovery proteomics (conducted on a TripleTOF 6600 mass spectrometer) followed by comparative quantitative MRM-MS analysis (conducted on a 6500+ QTRAP mass spectrometer)
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