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https://www.readbyqxmd.com/read/28324630/morphological-and-neurochemical-differences-in-peptidergic-nerve-fibres-of-the-mouse-vagina
#1
Christine Barry, Esther Ji, Harman Sharma, Lara Beukes, Pat Vilimas, Yvette DeGraaf, Dusan Matusica, Rainer V Haberberger
The vagina is innervated by a complex arrangement of sensory, sympathetic and parasympathetic nerve fibres that contain classical transmitters plus an array of neuropeptides and enzymes known to regulate diverse processes including blood flow and nociception. The neurochemical characteristics and distributions of peptide-containing nerves in the mouse vagina are unknown. This study used multiple labelling immunohistochemistry, confocal imaging and analysis to investigate the presence and colocalization of the peptides vasoactive intestinal polypeptide (VIP), calcitonin-gene related peptide (CGRP), substance P (SP), neuropeptide tyrosine (NPY) and the nitric oxide synthesizing enzyme neuronal nitric oxide synthase (nNOS) in nerve fibres of the murine vaginal wall...
March 21, 2017: Journal of Comparative Neurology
https://www.readbyqxmd.com/read/28324507/using-protein-fragment-complementation-assays-pca-and-peptide-arrays-to-study-telomeric-protein-protein-interactions
#2
Wenbin Ma, Ok-Hee Lee, Hyeung Kim, Zhou Songyang
Studying protein-protein interactions is critical to our understanding of signaling pathways. Telomere Interactome is assembled around telomeres and consists of proteins and factors from diverse pathways. Dissecting how this protein network contributes to telomere protection and length regulation requires the elucidation of the complex and dynamic interactions between the proteins within the interactome. Here, we focus on three assays, Bi-molecular Fluorescence Complementation (BiFC), Gaussia Luciferase Protein-fragment Complementation Assay (GLuc PCA), and OPAL peptide array, which have proven vital in our studies of telomere protein interaction networks...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28323576/tat-rasgap317-326-enhances-radiosensitivity-of-human-carcinoma-cell-lines-in-vitro-and-in-vivo-through-promotion-of-delayed-mitotic-cell-death
#3
Pelagia Tsoutsou, Alessandro Annibaldi, David Viertl, Jonathan Ollivier, Franz Buchegger, Marie-Catherine Vozenin, Jean Bourhis, Christian Widmann, Oscar Matzinger
The synthetic peptide TAT-RasGAP317-326 has been shown to potentiate the efficacy of anti-cancer drugs. In this study, we explored the action of TAT-RasGAP317-326 when combined with radiation by investigating its radiosensitizing activity in vitro and in vivo. To investigate the modulation of intrinsic radiosensitivity induced by TAT-RasGAP317-326, clonogenic assays were performed using four human cancer cell lines, HCT116 p53(+/+) (ATCC: CCL-247), HCT116 p53(-/-), PANC-1 (ATCC: CRL-1469) and HeLa (ATCC: CCL-2), as well as one nontumor cell line, HaCaT (CLS: 300493)...
March 21, 2017: Radiation Research
https://www.readbyqxmd.com/read/28322199/adrenomedullin-suppresses-vascular-endothelial-growth-factor-induced-vascular-hyperpermeability-and-inflammation-in-retinopathy
#4
Akira Imai, Yuichi Toriyama, Yasuhiro Iesato, Kazutaka Hirabayashi, Takayuki Sakurai, Akiko Kamiyoshi, Yuka Ichikawa-Shindo, Hisaka Kawate, Megumu Tanaka, Tian Liu, Xian Xian, Liuyu Zhai, Kun Dai, Keiya Tanimura, Teng Liu, Nanqi Cui, Akihiro Yamauchi, Toshinori Murata, Takayuki Shindo
Diabetic macular edema (DME) is caused by blood-retinal barrier breakdown associated with retinal vascular hyperpermeability and inflammation, and it is the major cause of visual dysfunction in diabetic retinopathy. Adrenomedullin (ADM) is an endogenous peptide first identified as a strong vasodilator. ADM is expressed in the eyes and is up-regulated in various eye diseases, although the pathophysiological significance is largely unknown. We investigated the effect of ADM on DME. In Kimba mice, which overexpress human vascular endothelial growth factor in their retinas, the capillary dropout, vascular leakage, and vascular fragility characteristic of diabetic retinopathy were observed, and these changes were accompanied by retinal inflammation...
March 16, 2017: American Journal of Pathology
https://www.readbyqxmd.com/read/28318230/label-free-discovery-array-platform-for-the-characterization-of-glycan-binding-proteins-and-glycoproteins
#5
Christopher John Gray, Antonio Sánchez-Ruiz, Ivana Šardzíková, Yassir A Ahmed, Rebecca Louise Miller, Juana E Reyes Martinez, Edward Pallister, Kun Huang, Peter Both, Mirja Hartmann, Hannah N Roberts, Robert Šardzik, Santanu Mandal, Jeremy E Turnbull, Claire E Eyers, Sabine L Flitsch
The identification of carbohydrate-protein interactions is central to our understanding of the roles of cell-surface carbohydrates (the glycocalyx), fundamental for cell-recognition events. Therefore there is a need for fast high-throughput biochemical tools to capture the complexity of these biological interactions. Here we describe a rapid method for qualitative label-free detection of carbohydrate-protein interactions on arrays of simple synthetic glycans, more com-plex natural glycosaminoglycans (GAG) and lectins/carbohydrate binding proteins using MALDI-ToF mass spectrometry...
March 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28316608/expression-of-16-nitrogenase-proteins-within-the-plant-mitochondrial-matrix
#6
Robert S Allen, Kimberley Tilbrook, Andrew C Warden, Peter C Campbell, Vivien Rolland, Surinder P Singh, Craig C Wood
The industrial production and use of nitrogenous fertilizer involves significant environmental and economic costs. Strategies to reduce fertilizer dependency are required to address the world's increasing demand for sustainable food, fibers, and biofuels. Biological nitrogen fixation, a process unique to diazatrophic bacteria, is catalyzed by the nitrogenase complex, and reconstituting this function in plant cells is an ambitious biotechnological strategy to reduce fertilizer use. Here we establish that the full array of biosynthetic and catalytic nitrogenase (Nif) proteins from the diazotroph Klebsiella pneumoniae can be individually expressed as mitochondrial targeting peptide (MTP)-Nif fusions in Nicotiana benthamiana...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28316235/evaluation-of-nanobody-conjugates-and-protein-fusions-as-bioanalytical-reagents
#7
Virginia J Bruce, Brian R McNaughton
Enzyme-linked immunosorbent assay (ELISA), flow cytometry, and Western blot are common bioanalytical techniques. Successful execution traditionally requires the use of one or more commercially available antibody-small-molecule dye, or antibody-reporter protein conjugates that recognize relatively short peptide tags (<15 amino acids). However, the size of antibodies, and their molecular complexity (by virtue of post-translational disulfide formation and glycosylation) typically requires either expression in mammalian cells or purification from immunized mammals...
March 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28315768/peptide-protein-interactions-within-human-hair-keratins
#8
Célia F Cruz, Teresa Matamá, Artur Cavaco-Paulo
We selected 1235 decapeptides from human hair proteins encoded by human genes of keratins and keratin associated proteins. The peptides were linked to glass arrays and screened for their affinity towards a solution of human hair extracted keratin fraction. Based on the physicochemical properties of the peptides, ten variables were studied: content of different types of amino acid side chains (cysteine, hydrophobic, polar, basic, acidic, aromatic rings, amide, alcohol side chains), isoelectric point, and net charge...
March 15, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28315359/antibodies-and-associates-partners-in-targeted-drug-delivery
#9
REVIEW
Patrick J Kennedy, Carla Oliveira, Pedro L Granja, Bruno Sarmento
Monoclonal antibodies (mAbs) are well established in the clinic due to their specificity and affinity to a diverse array of biochemical targets. More recently, mAbs are being exploited as targeting agents in modern drug delivery systems, aiming to bypass normal host tissue and to accumulate a therapeutic agent to a specific tissue or cell for enhanced pharmacology. At sizes ranging from ~10-100nm, antibody-based bioconjugates have opened up a whole new realm of clinical possibilities with several platforms emerging on the market...
March 14, 2017: Pharmacology & Therapeutics
https://www.readbyqxmd.com/read/28301141/reconstitution-and-substrate-specificity-of-the-radical-sam-thiazole-c-methyltransferase-in-thiomuracin-biosynthesis
#10
Nilkamal Mahanta, Zhengan Zhang, Graham A Hudson, Wilfred A van der Donk, Douglas A Mitchell
Thiomuracin is a thiopeptide antibiotic with potent activity towards Gram-positive drug-resistant bacteria. Thiomuracin is biosynthesized from a precursor peptide, TbtA, by a complex array of posttranslational modifications. One of several intriguing transformations is the C-methylation of thiazole, occurring at an unactivated sp2 carbon. Herein, we report the in vitro reconstitution of TbtI, the responsible radical S-adenosyl-methionine (rSAM) C-methyltransferase, which catalyzes the formation of 5-methylthiazole at a single site...
March 16, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28299740/detection-of-matrix-metalloproteinases-by-zymography
#11
Rajeev B Tajhya, Rutvik S Patel, Christine Beeton
Matrix metalloproteinases (MMPs) represent more than 20 zinc-containing endopeptidases that cleave internal peptide bonds, leading to protein degradation. They play a critical role in many physiological cell functions, including tissue remodeling, embryogenesis, and angiogenesis. They are also involved in the pathogenesis of a vast array of diseases, including but not limited to systemic inflammation, various cancers, and cardiovascular, neurological, and autoimmune diseases. Here, we describe gel zymography to detect MMPs in cell and tissue samples and in cell culture supernatants...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28299705/characterization-of-in-vivo-selected-bacteriophage-for-the-development-of-novel-tumor-targeting-agents-with-specific-pharmacokinetics-and-imaging-applications
#12
Jessica Newton-Northup, Susan L Deutscher
Bacteriophage (phage) display technology is a powerful strategy for the identification of peptide-based tumor targeting agents for drug discovery. Phage selections performed in vitro often result in many phage clones/peptides with similar properties and often similar sequence. However, these phage and corresponding peptides are selected, validated, and characterized outside the complicated milieu of a living animal. Thus, there is no guarantee that peptides from in vitro selections will successfully meet the requirements of an in vivo targeting compound...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28299684/bacterial-detection-using-peptide-based-platform-and-impedance-spectroscopy
#13
Hashem Etayash, Thomas Thundat, Kamaljit Kaur
Antimicrobial peptides have the ability to function as bio-recognition elements in the detection of bacteria. For instance, we showed that Leucocin A, an antimicrobial peptide from class IIa bacteriocins, binds gram-positive Listeria monocytogenes with higher affinity than other gram-positive bacteria like S. aureus, L. innocua, and E. faecalis. The binding was detected using impedance spectroscopy when Leucocin A immobilized on impedance electrodes binds bacteria from a sample. Here we highlight the strength of utilizing Leucocin A as a bio-recognition probe in biosensor platforms and provide details on its application in real-time bacterial detection using electrochemical impedance spectroscopy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28299594/dissecting-pistil-responses-to-incompatible-and-compatible-pollen-in-self-incompatibility-brassica-oleracea-using-comparative-proteomics
#14
Jing Zeng, Qiguo Gao, Songmei Shi, Xiaoping Lian, Richard Converse, Hecui Zhang, Xiaohong Yang, Xuesong Ren, Song Chen, Liquan Zhu
Angiosperms have developed self-incompatibility (SI) systems to reject self-pollen, thereby promoting outcrossing. The Brassicaceae belongs to typical sporophytic system, having a single S-locus controlled SI response, and was chosen as a model system to study SI-related intercellular signal transduction. In this regard, the downstream factor of EXO70A1 was unknown. Here, protein two-dimensional electrophoresis (2-DE) method and coupled with matrix-assisted laser desorption ionization/time of flight of flight mass spectrometry (MALDI-TOF -MS) and peptide mass fingerprinting (PMF) was used to further explore the mechanism of SI responses in Brassica oleracea L...
March 16, 2017: Protein Journal
https://www.readbyqxmd.com/read/28286864/in-vivo-expansion-of-melanoma-specific-t-cells-using-microneedle-arrays-coated-with-immune-polyelectrolyte-multilayers
#15
Qin Zeng, Joshua M Gammon, Lisa H Tostanoski, Yu-Chieh Chiu, Christopher M Jewell
Microneedles (MNs) are micron-scale polymeric or metallic structures that offer distinct advantages for vaccines by efficiently targeting skin-resident immune cells, eliminating injection-associated pain, and improving patient compliance. These advantages, along with recent studies showing therapeutic benefits achieved using traditional intradermal injections in human cancer patients, suggest MN delivery might enhance cancer vaccines and immunotherapies. We recently developed a new class of polyelectrolyte multilayers based on the self-assembly of model peptide antigens and molecular toll-like receptor agonists (TLRa) into ultrathin, conformal coatings...
February 13, 2017: ACS Biomaterials Science & Engineering
https://www.readbyqxmd.com/read/28284242/epstein-barr-virus-nuclear-antigen-1-interacts-with-regulator-of-chromosome-condensation-1-dynamically-throughout-the-cell-cycle
#16
Thibaut Deschamps, Quentin Bazot, Derek M Leske, Ruth MacLeod, Dimitri Mompelat, Lionel Tafforeau, Vincent Lotteau, Vincent Maréchal, George S Baillie, Henri Gruffat, Joanna B Wilson, Evelyne Manet
The Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is a sequence-specific DNA-binding protein that plays an essential role in viral episome replication and segregation, by recruiting the cellular complex of DNA replication onto the origin (oriP) and by tethering the viral DNA onto the mitotic chromosomes. Whereas the mechanisms of viral DNA replication are well documented, those involved in tethering EBNA1 to the cellular chromatin are far from being understood. Here, we have identified regulator of chromosome condensation 1 (RCC1) as a novel cellular partner for EBNA1...
February 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28282025/nuclear-export-receptor-crm1-recognizes-diverse-conformations-in-nuclear-export-signals
#17
Ho Yee Joyce Fung, Szu-Chin Fu, Yuh Min Chook
Nuclear export receptor CRM1 binds highly variable nuclear export signals (NESs) in hundreds of different cargoes. Previously we have shown that CRM1 binds NESs in both polypeptide orientations (Fung et al., 2015). Here, we show crystal structures of CRM1 bound to eight additional NESs which reveal diverse conformations that range from loop-like to all-helix, which occupy different extents of the invariant NES-binding groove. Analysis of all NES structures show 5-6 distinct backbone conformations where the only conserved secondary structural element is one turn of helix that binds the central portion of the CRM1 groove...
March 10, 2017: ELife
https://www.readbyqxmd.com/read/28273806/antimicrobial-dendrimeric-peptides-structure-activity-and-new-therapeutic-applications
#18
REVIEW
Mariano A Scorciapino, Ilaria Serra, Giorgia Manzo, Andrea C Rinaldi
Microbial resistance to conventional antibiotics is one of the most outstanding medical and scientific challenges of our times. Despite the recognised need for new anti-infective agents, however, very few new drugs have been brought to the market and to the clinic in the last three decades. This review highlights the properties of a new class of antibiotics, namely dendrimeric peptides. These intriguing novel compounds, generally made of multiple peptidic sequences linked to an inner branched core, display an array of antibacterial, antiviral and antifungal activities, usually coupled to low haemolytic activity...
March 3, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28258202/the-endocannabinoid-metabolite-prostaglandin-e2-pge2-glycerol-inhibits-human-neutrophil-functions-involvement-of-its-hydrolysis-into-pge2-and-ep-receptors
#19
Caroline Turcotte, Simona Zarini, Stéphanie Jean, Cyril Martin, Robert C Murphy, David Marsolais, Michel Laviolette, Marie-Renée Blanchet, Nicolas Flamand
The endocannabinoids 2-arachidonoyl-glycerol and N-arachidonoyl-ethanolamine mediate an array of pro- and anti-inflammatory effects. These effects are related, in part, to their metabolism by eicosanoid biosynthetic enzymes. For example, N-arachidonoyl-ethanolamine and 2-arachidonoyl-glycerol can be metabolized by cyclooxygenase-2 into PG-ethanolamide (PG-EA) and PG-glycerol (PG-G), respectively. Although PGE2 is a recognized suppressor of neutrophil functions, the impact of cyclooxygenase-derived endocannabinoids such as PGE2-EA or PGE2-G on neutrophils is unknown...
March 3, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28256805/targeted-label-free-quantification-of-interleukin-8-in-pma-activated-u937-cell-secretome-by-nanolc-esi-ms-ms-ssrm
#20
Pelin Esma Emirbayer, Kerstin F Gerer, Stefanie Hoyer, Monika Pischetsrieder
Monocytes are a part of the innate immune system. Their differentiation into macrophages changes their cellular proteome and secretome. Particularly secretome components such as cytokines are crucial for immune response and inflammation in many diseases. Differentiation of human lymphoma cell line U937 can be triggered by phorbol 12-myristate 13-acetate (PMA). Screening of the cytokine release in U937 upon PMA stimulation by cytometric bead array almost exclusively showed interleukin-8. Next, a label-free nanoLC-ESI-MS/MS-sSRM method for quantification of interleukin-8 in the cell secretome was established and applied to monitor the time kinetics of PMA treatment in different concentrations...
March 3, 2017: Proteomics
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