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Eliana Marzol, Cecilia Borassi, Mauro Bringas, Ana Sede, Diana Rosa Rodríguez Garcia, Luciana Capece, Jose M Estevez
Extensins (EXTs) are highly repetitive plant O-glycoproteins that require several post-translational modifications (PTMs) to become functional in plant cell walls. First, they are hydroxylated on contiguous proline residues; then, they are O-glycosylated on hydroxyproline (Hyp) and serine. After secretion into the apoplast, O-glycosylated EXTs form a tridimensional network organized by inter- and intra-Tyr linkages. Recent studies have made significant progress in the identification of the enzymatic machinery required to process EXTs, which includes prolyl 4-hydroxylases (P4Hs), glycosyltransferases (GTs), papain-type cysteine-endopeptidases (CEPs), and peroxidases (PERs)...
March 9, 2018: Molecular Plant
Ramanjeet Kaur, Santosh Kumar Tiwari
Bacteriocin LB44 was purified from cell-free supernatant (CFS) of Pediococcus pentosaceus LB44 using activity-guided chromatography techniques. It was stable up to 121 °C, pH 2.0-6.0, sensitive to proteinase K, papain and trypsin, and retained complete activity in the presence of organic solvents tested. The molecular weight of bacteriocin was ∼6 kDa and initial ten amino acid residues (GECGMCXECG) suggested a new compound. The loss in viable cell count and K+ ion efflux of target cells of Micrococcus luteus suggested bactericidal activity...
March 9, 2018: Biochemical and Biophysical Research Communications
Rita B Santos, Balakumaran Chandrasekar, Manoj K Mandal, Farnusch Kaschani, Markus Kaiser, Leonard Both, Renier Al van der Hoorn, Andreas Schiermeyer, Rita Abranches
Medicago truncatula is an established model for studying legume biology. More recently, it has also been exploited as a Molecular Farming platform for the production of recombinant proteins, with the successful expression of fungal and human proteins in plants and cell suspension cultures of this species. One of the challenges that now must be overcome is the degradation of final products during production and downstream processing stages. In the Medicago truncatula genome, there are more than 400 putative protease-encoding genes, but to date, the proteolytic content of Medicago cell cultures has not been studied...
March 12, 2018: Biotechnology Journal
Marc Planas, Martí Bernardo-Faura, Judith Katharina Paulus, Farnusch Kaschani, Markus Kaiser, Marc Valls, Renier van der Hoorn, Nuria Sanchez Coll
Activity-based protein profiling (ABPP) is a powerful proteomic technique to display protein activities in a proteome. It is based on the use of small molecular probes that react with the active site of proteins in an activity-dependent manner. We used ABPP to dissect the protein activity changes that occur in the intercellular spaces of tolerant (Hawaii 7996) and susceptible (Marmande) tomato plants in response to R. solanacearum, the causing agent of bacterial wilt, one of the most destructive bacterial diseases in plants...
March 9, 2018: Molecular & Cellular Proteomics: MCP
Tatiane Garcia Stancker, Stella Souza Vieira, Andrey Jorge Serra, Rafael do Nascimento Lima, Regiane Dos Santos Feliciano, José Antônio Silva, Solange Almeida Dos Santos, Marcia Ataize Dos Santos Vieira, Maíra Cecília Brandão Simões, Ernesto Cesar Leal-Junior, Paulo de Tarso Camillo de Carvalho
This study aimed to determine whether photobiomodulation therapy (PBMT) could improve the bioavailability and chondroprotective benefits of mesenchymal stem cells injected into the knees of rats used as an experimental model of osteoarthritis (OA) as well as reduce the expression of matrix metalloproteinases (MMPs) and degradation of type II collagen (COL2-1) in the cartilage. Adipose-derived stem/stromal cells (ADSCs) were collected from three male Fischer 344 rats and characterized by flow cytometry. Fifty female Fischer 344 rats were distributed into five groups of 10 animals each...
March 8, 2018: Lasers in Medical Science
Mariana N Machado, Flavia Mazzoli-Rocha, Natália V Casquilho, Tatiana Maron-Gutierrez, Victor H Ortenzi, Marcelo M Morales, Rodrigo S Fortunato, Walter A Zin
Murine papain-induced emphysema is a model that reproduces many of the features found in patients. Bone marrow-derived mononuclear cells (BMMC) have already been used to repair the alveolar epithelium in respiratory diseases, but not in the papain model. Thus, we hypothesized that BMMC could prevent the pathophysiological processes in papain-induced experimental emphysema. Female BALB/c mice received intratracheal instillation of 50 μL of saline (S groups) or papain (P groups, 10 IU/50 μl of saline) on days 1 and 7 of the experimental protocol...
2018: Frontiers in Physiology
Tomoyuki Odaka, Hiroaki Suetake, Tomoki Maeda, Toshiaki Miyadai
Recently, mammalian basophils have been highlighted as having roles in allergy and antiparasitic immunity; however, there is little information about the functions and evolutionary origin of basophils, because they are the least abundant leukocyte in most vertebrates. In this study, we characterized the teleost basophils that are abundant in the peripheral blood of fugu ( Takifugu rubripes ). Fugu basophils have two distinct granules: reddish-purple and dark violet ones. Teleost fish do not have IgG and IgE, but we found that fugu IgM bound on the surface of the basophils, and the cross-linked IgM induced degranulation of both types of granules...
March 7, 2018: Journal of Immunology: Official Journal of the American Association of Immunologists
Andrea B Stefanovic, Jelena R Jovanovic, Bojana D Balanc, Nataša Ž Šekuljica, Sonja M Jakovetic Tanaskovic, Marina B Dojcinovic, Zorica D Kneževic-Jugovic
The objective of this study was to discover the relationship between the ultrasound probe treatment (UPT) on egg white proteins (EWPs) before EWPs hydrolysis by different proteases, and the functional properties of the obtained hydrolysates. To fulfill this goal, the protein solubility, foaming, and emulsifying properties were studied as a function of the UPT time and then related to the surface characteristics and structural properties. The changes in the hydrolysates microstructures and macromolecular conformation, induced by the UPT, were followed using scanning electron microscope analyzis (SEM) and Fourier transforms infrared spectroscopy (FTIR)...
March 5, 2018: Poultry Science
Friederike Grosse-Holz, Luisa Madeira, Muhammad Awais Zahid, Molly Songer, Jiorgos Kourelis, Mary Fesenko, Sabrina Ninck, Farnusch Kaschani, Markus Kaiser, Renier A L van der Hoorn
Agroinfiltrated Nicotiana benthamiana is a flexible and scalable platform for recombinant protein (RP) production, but its great potential is hampered by plant proteases that degrade RPs. Here, we tested 29 candidate protease inhibitors (PIs) in agroinfiltrated N. benthamiana leaves for enhancing accumulation of three unrelated RPs: glycoenzyme α-Galactosidase; glycohormone erythropoietin (EPO); and IgG antibody VRC01. Of the previously described PIs enhancing RP accumulation, we found only cystatin SlCYS8 to be effective...
March 6, 2018: Plant Biotechnology Journal
Yujing Qin, Qingxia Yuan, Yuexing Zhang, Jialu Li, Xinjiao Zhu, Lingling Zhao, Jing Wen, Jikai Liu, Longyan Zhao, Jinhua Zhao
Enzyme-assisted extraction optimization, characterization and in vitro antioxidant activity of polysaccharides from sea cucumber Phyllophorus proteus (PPP) were investigated in the present study. The optimal extraction conditions with a yield of 6.44 ± 0.06% for PPP were determined as follows: Extraction time of 2.89 h, ratio of extraction solvent to raw material of 16.26 mL/g, extraction pH of 6.83, exraction temperature of 50 °C and papain concentration of 0.15%. Three purified fractions, PPP-1a, PPP-1b and PPP-2 with molecular weights of 369...
March 6, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
J Patel, S Lal, K Nuss, S P Wilshaw, B von Rechenberg, R M Hall, J L Tipper
Less than optimal particle isolation techniques have impeded analysis of orthopaedic wear debris in vivo. The purpose of this research was to develop and test an improved method for particle isolation from tissue. A volume of 0.018 mm3 of clinically relevant CoCrMo, Ti-6Al-4V or Si3 N4 particles was injected into rat stifle joints for seven days of in vivo exposure. Following sacrifice, particles were located within tissues using histology. The particles were recovered by enzymatic digestion of periarticular tissue with papain and proteinase K, followed by ultracentrifugation using a sodium polytungstate density gradient...
March 2, 2018: Acta Biomaterialia
Sachin Rustgi, Edouard Boex-Fontvieille, Christiane Reinbothe, Diter von Wettstein, Steffen Reinbothe
Plants have evolved an intricate regulatory network of proteases and corresponding protease inhibitors (PI), which operate in various biological pathways and serve diverse spatiotemporal functions during the sedentary life of a plant. Intricacy of the regulatory network can be anticipated from the observation that, depending on the developmental stage and environmental cue(s), either a single PI or multiple PIs regulate the activity of a given protease. On the other hand, the same PI often interacts with different targets at different places, necessitating another level of fine control to be added in planta...
2018: Communicative & Integrative Biology
Raman Pachaiappan, Ekant Tamboli, Aurovind Acharya, Chia-Hung Su, Subash C B Gopinath, Yeng Chen, Palaniyandi Velusamy
Enzyme hydrolysates (trypsin, papain, pepsin, α-chymotrypsin, and pepsin-pancreatin) of Tinospora cordifolia stem proteins were analyzed for antioxidant efficacy by measuring (1) 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging activity, (2) 2,20-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) radical scavenging capacity, and (3) Fe2+ chelation. Trypsin hydrolysate showed the strongest DPPH• scavenging, while α-chymotrypsin hydrolysate exhibited the highest ABTS+ scavenging and Fe2+ chelation...
2018: PloS One
Sebastian Ziemann, Karina van der Linde, Urs Lahrmann, Beyda Acar, Farnusch Kaschani, Tom Colby, Markus Kaiser, Yezhang Ding, Eric Schmelz, Alisa Huffaker, Nicholas Holton, Cyril Zipfel, Gunther Doehlemann
Localized control of cell death is crucial for the resistance of plants to pathogens. Papain-like cysteine proteases (PLCPs) regulate plant defence to drive cell death and protection against biotrophic pathogens. In maize (Zea mays), PLCPs are crucial in the orchestration of salicylic acid (SA)-dependent defence signalling. Despite this central role in immunity, it remains unknown how PLCPs are activated, and which downstream signals they induce to trigger plant immunity. Here, we discover an immune signalling peptide, Z...
February 26, 2018: Nature Plants
Mohd Faizan Siddiqui, Bilqees Bano
Carbendazim is a broad spectrum benzimidazole fungicide which is used to ensure plants' protection from pest and pathogens' invasion. The present work describes the impact of carbendazim (CAR) on garlic phytocystatin (GPC) which is a crucial plant regulatory protein. Interaction of carbendazim with GPC has been investigated through various biophysical techniques viz. UV absorption, fluorescence spectroscopy, isothermal titration calorimetry, far-UV circular dichroism and FTIR spectroscopy which showed binding between them with consequent modulatory effects...
February 2018: Pesticide Biochemistry and Physiology
J S Mendoza-Figueroa, A Kvarnheden, J Méndez-Lozano, E-A Rodríguez-Negrete, R Arreguín-Espinosa de Los Monteros, M Soriano-García
Tomato yellow leaf curl virus (TYLCV; genus Begomovirus; family Geminiviridae) infects mainly plants of the family Solanaceae, and the infection induces curling and chlorosis of leaves, dwarfing of the whole plant, and reduced fruit production. Alternatives for direct control of TYLCV and other geminiviruses have been reported, for example, the use of esterified whey proteins, peptide aptamer libraries or artificial zinc finger proteins. The two latter alternatives affect directly the replication of TYLCV as well as of other geminiviruses because the replication structures and sequences are highly conserved within this virus family...
February 2018: Pesticide Biochemistry and Physiology
Selvakumari Ulagesan, Kamatchi Sankaranarayanan, Amutha Kuppusamy
A peptide might be an exciting biomaterial or template for the development of novel wound-healing agents. In this report, it was isolated from the terrestrial snail Cryptozona bistrialis by enzymatic digestion and was evaluated for its in vitro wound-healing activity in NIH/3T3 mouse fibroblasts cell line and in vivo wound-healing activity in normal and diabetic-induced Wistar albino rats. The C. bistrialis protein was digested by the papain enzyme, and 21.79 kDa peptide (Cb-peptide) was purified by reversed-phase high-performance liquid chromatography and identified by MALDI (matrix-assisted laser desorption/ionization)-TOF analysis...
February 26, 2018: International Wound Journal
Mateusz Kędzior, Aleksandra Pawlak, Rafał Seredyński, Jacek Bania, Aleksandra Platt-Samoraj, Magdalena Czemplik, Elżbieta Klausa, Gabriela Bugla-Płoskońska, Jan Gutowicz
Cysteine proteases of the papain family, including mammalian cathepsins, play important physiological roles, however, their excessive activity may contribute to the development of various pathologies. Therefore, cysteine cathepsin inhibitors are being considered as promising drugs to treat cathepsin-driven diseases. Diverse saprophytic and parasitic microbes produce such inhibitors, which target the host's proteases playing pivotal roles in immune responses, thus leading to the survival of microbes within their host...
March 2018: Microbiological Research
Morteza Bahrami, Ahmad Homaei
Papain has a relatively small active site consisting of three residues: Cys-25, His-159, and Asn-175. The sulfhydryl group on Cys-25 often forms covalent bonds with substrates. His-159 supports Cys-25, and while Asn-175 does not directly participate in the catalytic mechanism, it keeps histidine-159 in its stabilized imidazole form. It is noticeable that sulfur atom of the catalytic Cys-25 is activated by cysteine through a SN 2-type reaction, but the mechanisms by which cysteine enhances papain activity remains unclear...
February 15, 2018: International Journal of Biological Macromolecules
Yan-Jie Gu, Min-Liang Zhu, Yan-Li Li, Chun-Hua Xiong
A new type of magnetic metal chelating carrier (PCMM-IDA-Cu2+ ) was prepared for the immobilization of papain, using chitosan as raw material, nano Fe3 O4 as magnetic material, SiO2 as porogen, iminodiacetic acid (IDA) as a chelating ligand, and binding with transition metal ion (Cu2+ ). The resulting products were well characterized by optical microscopy, scanning electron microscopy (SEM), vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA)...
February 15, 2018: International Journal of Biological Macromolecules
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