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Wei Zhou, Xin Wang, Dan Zhou, Yidan Ouyang, Jialing Yao
Increasing grain yield and improving grain quality are two important goals for rice breeding. A better understanding of the factors that contribute to the overall grain quantity and nutritional quality of rice will lay the foundation for developing new breeding strategies. RAG2 is a member of 14-16 kDa α-amylase/trypsin inhibitors in rice, which belong to the albumin of seed storage proteins. We found that RAG2 was specifically expressed in ripening seed and its transcription peak was between 14 and 21 days after flowering...
October 24, 2016: Plant Biotechnology Journal
Usama W Hawas, Radwan Al-Farawati, Lamia T Abou El-Kassem, Adnan J Turki
The endophytic fungus Fusarium equiseti was isolated from the brown alga Padina pavonica, collected from the Red Sea. The fungus was identified by its morphology and 18S rDNA. Cultivation of this fungal strain in biomalt-peptone medium led to isolation of 12 known metabolites of diketopeprazines and anthraquinones. The organic extract and isolated compounds were screened for their inhibition of hepatitis C virus NS3/4A protease (HCV PR). As a result, the fungal metabolites showed inhibition of HCV protease (IC50 from 19 to 77 μM), and the fungus was subjected to culture on Czapek's (Cz) media, with a yield of nine metabolites with potent HCV protease inhibition ranging from IC50 10 to 37 μM...
October 20, 2016: Marine Drugs
Noriaki Okimoto, Atsushi Suenaga, Makoto Taiji
In computational drug design, ranking a series of compound analogues in a manner that is consistent with experimental affinities remains a challenge. In this study, we evaluated the prediction of protein-ligand binding affinities using steered molecular dynamics simulations. First, we investigated the appropriate conditions for accurate predictions in these simulations. A conic harmonic restraint was applied to the system for efficient sampling of work values on the ligand unbinding pathway. We found that pulling velocity significantly influenced affinity predictions, but that the number of collectable trajectories was less influential...
October 24, 2016: Journal of Biomolecular Structure & Dynamics
Kartiga Natarajan, Koteswara R Gottipati, Kiflu Berhane, Buka Samten, Usha Pendurthi, Vijay Boggaram
BACKGROUND: Persistant inflammatory responses to infectious agents and other components in organic dust underlie lung injury and development of respiratory diseases. Organic dust components responsible for eliciting inflammation and the mechanisms by which they cause lung inflammation are not fully understood. We studied the mechanisms by which protease activities in poultry dust extracts and intracellular oxidant stress induce inflammatory gene expression in A549 and Beas2B lung epithelial cells...
October 22, 2016: Respiratory Research
Shobha Devi, Po-Hung Wu, Pei-Yu Chu, Yue-Pei Liu, Hsin-Lin Wu, Yen-Peng Ho
The impact of microwave irradiation on the in-solution digestion processes and the detection limit of proteins are systematically studied. Kinetic processes of many peptides produced through the trypsin digestion of various proteins under microwave heating at 50°C were investigated with MALDI-MS. This study also examines the detection limits and digestion completeness of individual proteins under microwave heating at 50°C and at different time intervals (1, 5 and 30 min) using LC-MS. We conclude that, if the peptides without missed cleavage dictate the detection limit, conventional digestion will lead to a better detection limit...
October 22, 2016: Electrophoresis
Gloria Isani, Enea Ferlizza, Aurora Cuoghi, Elisa Bellei, Emanuela Monari, Barbara Bianchin Butina, Carolina Castagnetti
Characterisation of the physiologic equine amniotic fluid (AF) proteome is a prerequisite to study its changes during diseases and discover new biomarkers. The aim of this study was to identify by a proteomic approach the most abundant proteins of equine AF. AF samples were collected at parturition from 24 healthy mares that delivered healthy foals. All samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on 4-12% gels. A pool of the 24 samples, after SDS-PAGE, was cut in 25 slices, trypsin-digested and analysed by mass spectrometry (MS) for protein identification...
October 11, 2016: Animal Reproduction Science
Shuyun Zhu, Lei He, Fang Zhang, Mengyuan Li, Shulin Jiao, Yan Li, Mengyuan Chen, Xian-En Zhao, Hua Wang
A highly sensitive fluorimetric assay has been developed for the evaluation of glutathione reductase (GR) activity and the screening of its inhibitors by using carbon quantum dots (CQDs) as the signal reporter. The detection mechanism is based on the following facts: (1) the fluorescence of CQDs can be quenched by Hg(II) through the strong CQDs-Hg(II) coordination; (2) GR can catalyze the reduction of oxidized glutathione (GSSG) into reduced glutathione (GSH), so that the fluorescence recovery of CQDs would take place resulting from the strong GSH-Hg(II) interaction; (3) GR can lose its catalytic reduction of GSSG in the presence of its inhibitors, which will inhibit the recovery of the quenched fluorescence of CQDs...
December 1, 2016: Talanta
Lufeng Zhang, Haiyan Qin, Wanwan Cui, Yang Zhou, Jianxiu Du
The development of new detection methods for proteases activity assay is important in clinical diagnostics and drug development. In this work, a simple, label-free, and turn-on fluorescent sensor was fabricated for trypsin, a protease produced in the pancreas. Cytochrome c, a natural substance of trypsin, could be selectively cleaved by trypsin into heme-peptide fragment. The produced heme-peptide fragment exhibited an intensive catalytic role on the H2O2-mediated the oxidation of thiamine to form strong fluorescent thiochrome...
December 1, 2016: Talanta
Caitlin D Allen, A James Link
Lasso peptides exist naturally in a threaded state as [1]rotaxanes, and we reasoned that lasso peptides cleaved in their loop region could serve as building blocks for catenanes. Mutagenesis of the lasso peptide microcin J25 (MccJ25) with two cysteine residues followed by cleavage of the peptide with trypsin led to a [2]rotaxane structure that self-assembled into a [3]catenane and [4]catenanes at room temperature in aqueous solution. The [3]catenane represents the smallest ring size of a catenane composed solely of polypeptide segments...
October 21, 2016: Journal of the American Chemical Society
Blake T Riley, Olga Ilyichova, Mauricio G S Costa, Benjamin T Porebski, Simon J de Veer, Joakim E Swedberg, Itamar Kass, Jonathan M Harris, David E Hoke, Ashley M Buckle
The kallikrein-related peptidase (KLK) family of proteases is involved in many aspects of human health and disease. One member of this family, KLK4, has been implicated in cancer development and metastasis. Understanding mechanisms of inactivation are critical to developing selective KLK4 inhibitors. We have determined the X-ray crystal structures of KLK4 in complex with both sunflower trypsin inhibitor-1 (SFTI-1) and a rationally designed SFTI-1 derivative to atomic (~1 Å) resolution, as well as with bound nickel...
October 21, 2016: Scientific Reports
Intidhar Bkhairia, Rabeb Ben Slama Ben Salem, Rim Nasri, Mourad Jridi, Sofiane Ghorbel, Moncef Nasri
Bioactive Liza aurata protein hydrolysates (LAPHs) were prepared by treatment with Trypsine (PH-TR), Esperase (PH-ES), enzyme preparations from Pseudomonas aeruginosa A2 (PH-A2), Bacillus subtilis A26 (PH-A26) and Liza aurata (PH-LA). Their functional properties and antioxidant activities were evaluated. The hydrolysates had degree of hydrolysis (DH) values ranging from 8.15 % to 13.05 %. Reverse-phase HPLC analyses of the LAPHs showed considerable variation in peptide composition. All hydrolysates had high protein content (83...
July 2016: Journal of Food Science and Technology
M R Williams, T Nakatsuji, J A Sanford, A F Vrbanac, R L Gallo
Bacteria that reside on the skin can influence the behavior of the cutaneous immune system, but the mechanisms responsible for these effects are incompletely understood. Colonization of the skin by Staphylococcus aureus (S. aureus) is increased in atopic dermatitis (AD) and can result in increased severity of the disease. In this study we show that S. aureus stimulates human keratinocytes to increase their endogenous protease activity, including specific increases in trypsin activity. This increased protease activity coincided with increased expression of mRNA for kallikreins (KLKs), with KLK6, 13, and 14 showing the greatest induction after exposure to S...
October 17, 2016: Journal of Investigative Dermatology
Monika Pietrowska, Hanna C Diehl, Grzegorz Mrukwa, Magdalena Kalinowska-Herok, Marta Gawin, Mykola Chekan, Julian Elm, Grzegorz Drazek, Anna Krawczyk, Dariusz Lange, Helmut E Meyer, Joanna Polanska, Corinna Henkel, Piotr Widlak
Determination of the specific type of thyroid cancer is crucial for the prognosis and selection of treatment of this malignancy. However, in some cases appropriate classification is not possible based on histopathological features only, and it might be supported by molecular biomarkers. Here we aimed to characterize molecular profiles of different thyroid malignancies using mass spectrometry imaging (MSI) which enables the direct annotation of molecular features with morphological pictures of an analyzed tissue...
October 17, 2016: Biochimica et Biophysica Acta
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
Wimonrat Panpetch, Jennifer K Spinler, James Versalovic, Somying Tumwasorn
BACKGROUND: Interleukin (IL)-8 is the key agent for initiating an inflammatory response to infection with Helicobacter pylori. Some strains of Lactobacillus spp. are known to colonize the stomach and suppress inflammation caused by H. pylori. In this study, we characterized two gastric-derived lactobacilli, Lactobacillus salivarius (LS) strains B37 and B60, capable of inhibiting H. pylori-induced IL-8 production by gastric epithelial cells. RESULTS: Conditioned media from LS-B37 and LS-B60 suppressed H...
October 18, 2016: BMC Microbiology
Viola Grabs, Anna Kersten, Bernhard Haller, Siegmund Braun, David C Nieman, Martin Halle, Johannes Scherr
INTRODUCTION: Vigorous and prolonged exercise such as marathon running increases inflammatory markers and the risk of upper respiratory illness (URI) in athletes. Nutritional supplements are being tested as countermeasures of exercise-induced inflammation and immune dysfunction. METHODS: In this prospective randomized, double-blind, placebo-controlled Phase I trial, healthy male runners (N = 138, age 42 ± 11 y) were supplemented with rutoside (600-1200 mg/day) and hydrolytic enzymes (540-1,080 mg/day bromelain, 288-576 mg/day trypsin) (WOB) or placebo (PL) for one week before and two weeks after the Munich Marathon 2013...
October 6, 2016: Medicine and Science in Sports and Exercise
Ahmad Bani-Jaber, Iyad Alshawabkeh, Samaa Abdullah, Imad Hamdan, Adel Ardakani, Maha Habash
Due to its unique properties, such as biodegradability, biocompatibility, high amphiphilic property, and micelle formation, casein (CS) has been increasingly studied for drug delivery. We used CS as a drug carrier in solid dispersions (SDs) and evaluated the effect of its degradation by trypsin on drug dissolution from the dispersions. SDs of CS and mefenamic acid (MA) were prepared by physical mixing, kneading, and coprecipitation methods. In comparison to pure MA, the dispersions were evaluated for drug-protein interaction, loss of drug crystalinity, and drug morphology by differential scanning calorimetry, X-ray diffractometry, Fourier transform infrared spectroscopy, and scanning electron microscopy...
October 17, 2016: AAPS PharmSciTech
Ulvi K Gursoy, Mervi Gursoy, Eija Könönen, Herman O Sintim, Veli-Jukka Uitto, Stina Syrjänen
In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an air-liquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model...
October 17, 2016: Cytotechnology
Ji-Hua Yu, Yang-Yang Li, Mian Xiang, Jian-Quan Zhu, Xin-He Huang, Wan-Jun Wang, Rui Tan, Jia-Yu Zhou, Hai Liao
OBJECTIVES: To clone and characterize a novel bi-functional α-amylase/subtilisin inhibitor (LASI) from the rhizome of Ligusticum chuanxiong, a traditional Chinese medicine. RESULTS: The LASI showed strong homology with members of the Kunitz trypsin inhibitor family. Its putative amino acid sequence has a 40 % identity with that of the α-amylase/subtilisin inhibitor from rice. LASI gene without signal peptide was expressed in E. coli Rosetta. After purification, the recombinant LASI protein was inhibitory against not only α-amylase from porcine pancreas, Helicoverpa armigera, Spodoptera litura and Plutella xylostella, but also subtilisin A, but not against trypsin or chymotrypsin...
October 17, 2016: Biotechnology Letters
Keiji Matsumoto, Yoshitaka Suetsugu, Yoshiaki Tanaka, Toyomi Kotaki, Shin G Goto, Tetsuro Shinoda, Sakiko Shiga
Juvenile hormone (JH) biosynthesis is inhibited under short-day conditions in the brown-winged green bug Plautia stali. We investigated allatostatic molecules in the brain of P. stali. Methanol brain extracts strongly inhibited JH biosynthesis. The allatostatic activities of the brain extracts were heat stable but gently suppressed by trypsin treatment, indicating that the allatostatic molecules were peptides. Grybi-MIP1, found in Gryllus bimaculatus as an allatostatic molecule, inhibited JH biosynthesis in P...
October 14, 2016: Journal of Insect Physiology
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