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Protein digestion

Donatella Nardiello, Clelia Prattichizzo, Maria Teresa Rocchetti, Loreto Gesualdo, Diego Centonze
A dedicated proteomic approach based on nano-Liquid Chromatography coupled with tandem mass spectrometry in ion trap is proposed for the analysis of proteins trapped in sorbent resin cartridges used to remove inflammatory mediators from blood by coupled plasma filtration adsorption (CPFA). The final purpose of the proposed proteomic approach was to obtain a reference map of plasma proteins trapped in CPFA sorbents used for the extracorporeal blood purification of healthy pigs, with the potential impact to design new bio-filters able to control the inflammatory imbalance under pathological conditions, such as severe sepsis...
October 12, 2016: Journal of Pharmaceutical and Biomedical Analysis
Yan Long, Wen-Bin Tsai, Jeffrey T Chang, Marcos Estecio, Medhi Wangpaichitr, Naramol Savaraj, Lynn G Feun, Helen H W Chen, Macus Tien Kuo
Many human tumors require extracellular arginine (Arg) for growth because the key enzyme for de novo biosynthesis of Arg, argininosuccinate synthetase 1 (ASS1), is silenced. These tumors are sensitive to Arg-starvation therapy using pegylated arginine deiminase (ADI-PEG20) which digests extracellular Arg. Many previous studies reported that ASS1 silencing is due to epigenetic inactivation of ASS1 expression by DNA methylation, and that the demethylation agent 5-aza-deoxycytidine (Aza-dC) can induce ASS1 expression...
September 28, 2016: Oncotarget
N C N Perera, G I Godahewa, Jehee Lee
Mitogen-activated protein kinase (MAPK) is involved in the regulation of cellular events by mediating signal transduction pathways. MAPK1 is a member of the extracellular-signal regulated kinases (ERKs), playing roles in cell proliferation, differentiation, and development. This is mainly in response to growth factors, mitogens, and many environmental stresses. In the current study, we have characterized the structural features of a homolog of MAPK1 from disk abalone (AbMAPK1). Further, we have unraveled its expressional kinetics against different experimental pathogenic infections or related chemical stimulants...
October 17, 2016: Fish & Shellfish Immunology
Hongjun Xia, Guangping Wan, Junlong Zhao, Jiawei Liu, Quan Bai
High performance liquid chromatography (HPLC) is a kind of efficient separation technology and has been used widely in many fields. Micro-sized porous silica microspheres as the most popular matrix have been used for fast separation and analysis in HPLC. In this paper, the monodisperse large-porous silica microspheres with controllable size and structure were successfully synthesized with polymer microspheres as the templates and characterized. First, the poly(glycidyl methacrylate-co-ethyleneglycol dimethacrylate) microspheres (PGMA-EDMA) were functionalized with tetraethylenepentamine (TEPA) to generate amino groups which act as a catalyst in hydrolysis of tetraethyl orthosilicate (TEOS) to form Si-containing low molecular weight species...
October 13, 2016: Journal of Chromatography. A
Jing Chen, Sudhirkumar Shinde, Prabal Subedi, Celina Wierzbicka, Börje Sellergren, Stefan Helling, Katrin Marcus
Selective enrichment techniques are essential for mapping of protein posttranslational modifications (PTMs). Phosphorylation is one of the PTMs which continues to be associated with significant analytical challenges. Particularly problematic are tyrosine-phosphorylated peptides (pY-peptides) resulting from tryptic digestion which commonly escape current chemo- or immuno- affinity enrichments and hence remain undetected. We here report on significant improvements in this regard using pY selective molecularly imprinted polymers (pY-MIPs)...
October 11, 2016: Journal of Chromatography. A
Saehun Mun, Jisu Kim, David Julian McClements, Yong-Ro Kim, Yongdoo Choi
A whey protein isolate-rhodamine B conjugate (WPI-RB) was synthesized to visualize changes in the location of a protein emulsifier in oil-in-water emulsions during digestion. An oil-soluble dye (Nile Red) was used to visualize changes in the lipid phase during digestion. Protein-labeled and lipid-labeled emulsions were passed through a simulated gastrointestinal tract consisting of mouth, stomach, and intestinal phases, and changes in protein and lipid location and morphology were monitored using confocal laser scanning microscopy...
March 15, 2017: Food Chemistry
Wyller Max Ferreira Silva, Bárbara Biduski, Karina Oliveira Lima, Vânia Zanella Pinto, Jéssica Fernanda Hoffmann, Nathan Levien Vanier, Alvaro Renato Guerra Dias
The objective of this study was to evaluate the effects of the heat-moisture treatment (HMT) applied to paddy rice grains on the physicochemical properties, in vitro starch digestibility, and molecular weight distribution of proteins in rice flour. The paddy rice grains were adjusted to 13%, 16%, and 18% moisture and autoclaved at 121°C for 30 and 60min. The HMT promoted a reduction of the amylose content, the swelling power, and the solubility of the rice flour. Changes in the relative crystallinity and molecular weight of the proteins extracted with a sodium phosphate buffer containing 2...
March 15, 2017: Food Chemistry
Laura Acevedo-Pacheco, Sergio O Serna-Saldívar
BACKGROUND: One way to diminish protein malnutrition in children is by enriching cereal-based flours for the manufacturing of maize tortillas, wheat flour tortillas, and yeast-leavened breads, which are widely consumed among low socio-economic groups. OBJECTIVE: The aim was to determine and compare the essential amino acid (EAA) scores, protein digestibility corrected amino acid scores (PDCAAS), and in vivo protein quality (protein digestibility, protein efficiency ratio (PER), biological values (BV), and net protein utilization (NPU) values) of regular versus soybean-fortified maize tortillas, yeast-leavened bread, and wheat flour tortillas...
2016: Food & Nutrition Research
C O Brito, J L L Dutra, T N Dias, L T Barbosa, C S Nascimento, A P G Pinto, L F T Albino, R P M Fernandes, M S Macário, J S Melo
The aim of this study was to evaluate the effect of dietary lysine on performance, protein deposition and respiratory chain gene expression in male broilers. A total of 252 Cobb 500 broilers were distributed, in a completely randomized design, into four treatments with seven replicates of nine birds per experimental unit. Experimental treatments consisted of diets based on corn and soybean meal, with four levels of digestible lysine: 1.016%, 1.099%, 1.182% and 1.265%. The increase in the level of digestible lysine in the diet provided higher weight gains, feed efficiency and body protein deposition...
October 21, 2016: Animal: An International Journal of Animal Bioscience
Jingni Wu, Yiming Wang, Sook-Young Park, Sang Gon Kim, Ju Soon Yoo, Sangryeol Park, Ravi Gupta, Kyu Young Kang, Sun Tae Kim
Rice blast disease caused by Magnaporthe oryzae is one of the most devastating fungal diseases of rice and results in a huge loss of rice productivity worldwide. During the infection process, M. oryzae secretes a large number of glycosyl hydrolase proteins into the host apoplast to digest the cell wall and facilitate fungal ingression into host tissues. In this study, we identified a novel arabinofuranosidase-B (MoAbfB) protein that is secreted by M. oryzae during fungal infection. Deletion of MoAbfB from M...
2016: PloS One
Ying Zhang, Xi Wang, Dan Cui, Jun Zhu
The human whole saliva is a vital body fluid to study the physiology and pathology of oral cavity. As a powerful technique for biomarker discovery, mass spectrometry-based proteomic strategy has been introduced to saliva analysis and identified hundreds of proteins and N-glycosylation sites. However, there still lacks of quantitative analysis, which is necessary for biomarker screening and biological researches. In this study, we established an integrated workflow by the combination of stable isotope dimethyl labeling, HILIC enrichment and high resolution mass spectrometry for both quantification of global proteome and N-glycoproteome of human saliva from oral ulcer patients...
October 20, 2016: Proteomics
Liping Sun, Weidan Chang, Qingyu Ma, Yongliang Zhuang
In this study, the stable collagen hydrolysate was prepared by alcalase hydrolysis and twice simulated gastrointestinal digestion from Alaska pollock skin. The characteristics of hydrolysates and antioxidant activities in vitro, including 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS(•+)) scavenging activity, ferric-reducing antioxidant power (FRAP) and hydroxyl radical (OH·) scavenging activity, were determined. After twice simulated gastrointestinal digestion of skin collagen (SGI-2), the degree of hydrolysis (DH) reached 26...
October 17, 2016: Marine Drugs
L Chen, G Guo, X J Yuan, J Zhang, A Y Wen, X H Sun, T Shao
The objective of this study was to determine the effect of ensiling different ratios of whole crop oat to lucerne on fermentation quality, aerobic stability and in vitro digestibility of silage on the Tibetan plateau. Four experimental treatments were produced varying in the ratio of forages on a fresh matter (FM) basis: 1) 100% oat (control, dry matter (DM) content: 317 g/kg), 2) 90% oat + 10% lucerne (OL10, DM content: 316 g/kg), 3) 80% oat+ 20% lucerne (OL20, DM content: 317 g/kg) and 4) 70% oat+ 30% lucerne (OL30, DM content: 318 g/kg)...
October 20, 2016: Journal of Animal Physiology and Animal Nutrition
Martin Wieczorek, Karl-Heinz Gührs, Thorsten Heinzel
Posttranslational acetylation of lysine residues has been discovered as multifaceted regulatory modification for various nuclear, cytoplasmic, and mitochondrial proteins. The implementation of high-resolution and high-throughput mass spectrometry (MS) approaches has led to the identification of a hitherto underappreciated, large number of acetylation sites for a broad spectrum of cellular proteins. In this chapter, we describe a comprehensive protocol for the purification of an in vivo-acetylated, ectopically expressed, FLAG-epitope tagged nonhistone protein through immunoprecipitation (IP)...
2017: Methods in Molecular Biology
Martin Jopcik, Jana Moravcikova, Ildiko Matusikova, Miroslav Bauer, Miroslav Rajninec, Jana Libantova
Chitinase gene from the carnivorous plant, Drosera rotundifolia , was cloned and functionally characterised. Plant chitinases are believed to play an important role in the developmental and physiological processes and in responses to biotic and abiotic stress. In addition, there is growing evidence that carnivorous plants can use them to digest insect prey. In this study, a full-length genomic clone consisting of the 1665-bp chitinase gene (gDrChit) and adjacent promoter region of the 698 bp in length were isolated from Drosera rotundifolia L...
October 19, 2016: Planta
Kristin Surmann, Marjolaine Simon, Petra Hildebrandt, Henrike Pförtner, Stephan Michalik, Vishnu M Dhople, Barbara M Bröker, Frank Schmidt, Uwe Völker
To simultaneously obtain proteome data of host and pathogen from an internalization experiment, human alveolar epithelial A549 cells were infected with Staphylococcus aureus HG001 which carried a plasmid (pMV158GFP) encoding a continuously expressed green fluorescent protein (GFP). Samples were taken hourly between 1.5 h and 6.5 h post infection. By fluorescence activated cell sorting GFP-expressing bacteria could be enriched from host cell debris, but also infected host cells could be separated from those which did not carry bacteria after contact (exposed)...
June 2016: Data in Brief
Bruno Tilocca, Maren Witzig, Markus Rodehutscord, Jana Seifert
The chicken gastrointestinal tract (GIT) harbours a complex microbial community, involved in several physiological processes such as host immunomodulation and feed digestion. For the first time, the present study analysed dietary effects on the protein inventory of the microbiome in crop and ceca of broilers. We performed quantitative label-free metaproteomics by using 1-D-gel electrophoresis coupled with LC-MS/MS to identify the structural and functional changes triggered by diets supplied with varying amount of mineral phosphorous (P) and microbial phytase (MP)...
2016: PloS One
Ronaldo Sousa Oliveira Filho, Lia Mara Kauchi Ribeiro, Lucia Caruso, Patricia Azevedo Lima, Náglia Raquel Teixeira Damasceno, Francisco García Soriano
INTRODUCTION: Quality Indicators for Nutritional Therapy (QINT) allow a practical assessment of nutritional therapy (NT) quality. OBJECTIVE: To apply and monitor QINT for critically ill patients at nutritional risk. METHODS: Cross sectional study including critically ill patients > 18 years old, at nutritional risk, on exclusive enteral (ENT) or parenteral nutritional therapy (PNT) for > 72 hours. After three consecutive years, 9 QINT were applied and monitored...
September 20, 2016: Nutrición Hospitalaria: Organo Oficial de la Sociedad Española de Nutrición Parenteral y Enteral
Fernando J Sialana, Peter Gulyassy, Peter Májek, Evelina Sjöstedt, Viktor Kis, André C Müller, Elena L Rudashevskaya, Jan Mulder, Keiryn L Bennett, Gert Lubec
The molecular composition of synaptic signal transduction machineries shapes synaptic neurotransmission. The repertoire of receptors, transporters and channels (RTCs) comprises major signaling events in the brain. RTCs are conventionally studied by candidate immunohistochemistry and biochemistry, which are low throughput with resolution greatly affected by available immunoreagents and membrane interference. Therefore, a comprehensive resource of synaptic brain RTCs is still lacking. In particular, studies on the detergent-soluble synaptosomal fraction, known to contain transporters and channels, are limited...
October 19, 2016: Proteomics
Laura Bouché, Maria Panico, Paul Hitchen, Daniel Binet, Federico Sastre, Alexandra Faulds-Pain, Esmeralda Valiente, Evgeny Vinogradov, Annie Aubry, Kelly Fulton, Susan Twine, Susan M Logan, Brendan W Wren, Anne Dell, Howard R Morris
Glycosylation of flagellins is a well recognized property of many bacterial species. In this study we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile. We used mass spectrometry (nano LC- MS and MS/MS analysis) to identify a number of putative glycopeptides which carried a variety of glycoform substitutions each of which was linked through an initial HexNAc residue to Ser or Thr. Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity...
October 7, 2016: Journal of Biological Chemistry
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