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https://www.readbyqxmd.com/read/29786787/protocols-for-migration-and-invasion-studies-in-prostate-cancer
#1
Arjanneke F van de Merbel, Geertje van der Horst, Jeroen T Buijs, Gabri van der Pluijm
Prostate cancer is the most common malignancy diagnosed in men in the western world. The development of distant metastases and therapy resistance are major clinical problems in the management of prostate cancer patients. In order for prostate cancer to metastasize to distant sites in the human body, prostate cancer cells have to migrate and invade neighboring tissue. Cancer cells can acquire a migratory and invasive phenotype in several ways, including single cell and collective migration. As a requisite for migration, epithelial prostate cancer cells often need to acquire a motile, mesenchymal-like phenotype...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29784297/destabilization-of-uht-milk-by-protease-aprx-from-pseudomonas-fluorescens-and-plasmin
#2
Chunyue Zhang, Etske Bijl, Kasper Hettinga
Destabilization of UHT milk during its shelf life is mainly promoted by the residual proteolytic activity attributed to the psychrotrophic bacterial proteases and native milk proteases. In this study, we built skim UHT milk-based model systems to which either the major bacterial protease (AprX from Pseudomonas fluorescens), or the major native milk protease (plasmin) was added, to allow a direct comparison between the destabilization of skim UHT milk by both categories of enzymes. The physical and chemical properties were studied during 6 weeks...
October 15, 2018: Food Chemistry
https://www.readbyqxmd.com/read/29783071/mef2a-regulates-calpain-3-expression-in-l6-myoblasts
#3
Ronghua Wu, Jun Wang, Jian Yao, Zhangji Dong, Yan Liu, Mei Liu
Calpain 3 (Capn3), a skeletal muscle-specific member of the calpain family, executes some non-proteolytic functions besides its role as a Ca2+ -regulated proteolytic enzyme. Previously, we found that changes in Capn3 expression were linearly correlated with the degree of muscular atrophy following reversible sciatic nerve injury and that knockdown of Capn3 gene expression promoted myoblast differentiation. While the regulation of capn3 gene expression is interesting, transcriptional regulation of Capn3 is still unclear...
May 18, 2018: Gene
https://www.readbyqxmd.com/read/29778758/major-traits-of-the-senescent-phenotype-of-nucleus-pulposus-intervertebral-disc-cells-persist-under-the-specific-microenvironmental-conditions-of-the-tissue
#4
Anastasios Kouroumalis, Eleni Mavrogonatou, Olga D Savvidou, Panayiotis J Papagelopoulos, Harris Pratsinis, Dimitris Kletsas
Intervertebral discs (IVDs) are the joints of the spine, mainly consisting of extracellular matrix (ECM) with a low number of cells embedded therein. Low cellularity stems from nutrient deprivation due to the lack of blood supply, as well as from the hypoxic and hyperosmotic conditions prevailing in the tissue. Intervertebral disc degeneration (IDD) has been firmly connected with low back pain, a major age-related disease, whereas degenerated discs have been characterized by increased proteolytic activity and accumulation of senescent cells...
May 17, 2018: Mechanisms of Ageing and Development
https://www.readbyqxmd.com/read/29775750/the-effects-of-capn1-gene-inactivation-on-the-differential-expression-of-genes-in-skeletal-muscle
#5
William T Olive, Brittney N Keel, Amanda K Lindholm-Perry, Justyna Horodyska, Andrew P Foote
Protein turnover is required for muscle growth and regeneration and several proteolytic enzymes, including the calpains, degrade myofibrillar proteins during this process. In a previous experiment, phenotypic differences were observed between μ-calpain knockout (KO) and wild type (WT) mice, including nutrient accretion and fiber type differences. These changes were particularly evident as the animals aged. Thus, we utilized 18 mice (9 KO and 9 WT) to compare transcript abundance to identify differentially expressed genes (DEGs) at 52 wk of age...
May 15, 2018: Gene
https://www.readbyqxmd.com/read/29773673/lysosomal-proteome-and-secretome-analysis-identifies-missorted-enzymes-and-their-non-degraded-substrates-in-mucolipidosis-iii-mouse-cells
#6
Giorgia Di Lorenzo, Renata Voltolini Velho, Dominic Winter, Melanie Thelen, Shiva Ahmadi, Michaela Schweizer, Raffaella De Pace, Kerstin Cornils, Timur Alexander Yorgan, Saskia Grüb, Irm Hermans-Borgmeyer, Thorsten Schinke, Sven Müller-Loennies, Thomas Braulke, Sandra Pohl
Targeting of soluble lysosomal enzymes requires mannose 6-phosphate (M6P) signals whose formation is initiated by the hexameric N-acetylglucosamine (GlcNAc)-1-phosphotransferase complex (α2β2γ2). Upon proteolytic cleavage by site-1 protease, the α/β-subunit precursor is catalytically activated but the functions of γ-subunits (Gnptg) in M6P modification of lysosomal enzymes are unknown. To investigate this, we analyzed the Gnptg expression in mouse tissues, primary cultured cells, and in Gnptg reporter mice in vivo, and found high amounts in the brain, eye, kidney, femur, vertebra and fibroblasts...
May 17, 2018: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/29771464/degradation-of-amyloid-%C3%AE-peptide-by-neprilysin-expressed-from-borna-disease-virus-vector
#7
Madoka Sakai, Sakiho Ueda, Takuji Daito, Megumi Asada-Utsugi, Yumiko Komatsu, Ayae Kinoshita, Takakuni Maki, Akira Kuzuya, Ryosuke Takahashi, Akiko Makino, Keizo Tomonaga
Accumulation of amyloid β (Aβ40 and Aβ42) in the brain is a characteristic of Alzheimer's disease (AD). Because neprilysin (NEP) is a major Aβ-degrading enzyme, NEP delivery in the brain is a promising gene therapy for AD. Borna disease virus (BoDV) vector enables long-term transduction of foreign genes in the central nerve system. Here, we evaluated the proteolytic ability of NEP transduced by the BoDV vector and found that the amounts of Aβ40 and Aβ42 significantly decreased, which suggests that NEP expressed from the BoDV vector is functional to degrade Aβ...
May 17, 2018: Microbiology and Immunology
https://www.readbyqxmd.com/read/29771001/total-chemical-synthesis-of-sumo-and-sumo-abps-to-profile-activity-of-sumo-specific-proteases
#8
Monique Priscilla Catharina Mulder, Remco Merkx, Katharina Witting, Dharjath Hameed, Dris El Atmioui, Lindsey Lelieveld, Frauke Liebelt, Jacques Neefjes, Ilana Berlin, Alfred Vertegaal, Huib Ovaa
SUMO, a post-translational modifier critical for cell cycle progression and genome stability, plays a role in tumorigenesis rendering SUMO-specific enzymes as potential pharmacological targets. However, systematic generation of tools for activity profiling of SUMO-specific enzymes has proven challenging, leading us to develop a diversifiable synthetic platform for a SUMO toolbox using a direct linear synthesis protocol, permitting N- and C-terminal labeling strategies to incorporate dyes and reactive warheads, respectively...
May 17, 2018: Angewandte Chemie
https://www.readbyqxmd.com/read/29768467/alpha-7-nicotinic-receptors-attenuate-neurite-development-through-calcium-activation-of-calpain-at-the-growth-cone
#9
Justin R King, Nadine Kabbani
The α7 nicotinic acetylcholine receptor (nAChR) is a ligand-gated ion channel that plays an important role in cellular calcium signaling contributing to synaptic development and plasticity, and is a key drug target for the treatment of neurodegenerative conditions such as Alzheimer's disease. Here we show that α7 nAChR mediated calcium signals in differentiating PC12 cells activate the proteolytic enzyme calpain leading to spectrin breakdown, microtubule retraction, and attenuation in neurite growth. Imaging in growth cones confirms that α7 activation decreases EB3 comet motility in a calcium dependent manner as demonstrated by the ability of α7 nAChR, ryanodine, or IP3 receptor antagonists to block the effect of α7 nAChR on growth...
2018: PloS One
https://www.readbyqxmd.com/read/29767556/hydronephrosis-is-associated-with-elevated-plasmin-in-urine-in-pediatric-patients-and-rats-and-changes-in-ncc-and-%C3%AE-enac-abundance-in-rat-kidney
#10
Rikke Zachar, Ammar Al-Mashhadi, Henrik Dimke, Per Svenningsen, Boye L Jensen, Mattias Carlström
Obstruction of urine flow at the level of the pelvo-ureteric junction (UPJO) and subsequent development of hydronephrosis is one of the most common congenital renal malformations. UPJO is associated with development of salt-sensitive hypertension, which is set by the obstructed kidney, and with a stimulated renin-angiotensin-aldosterone system (RAAS) in rodent models. This study aimed at investigating the hypothesis that i) in pediatric patients with UPJO the RAAS is activated prior to surgical relief of the obstruction; ii) in rats with UPJO the RAAS activation is reflected by increased abundance of renal aldosterone-stimulated Na+ transporters; and iii) the injured UPJO kidney allows aberrant filtration of plasminogen leading to proteolytic activation of the epithelial sodium channel gamma subunit (γ-ENaC)...
May 16, 2018: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/29767335/regulation-of-enzyme-activities-in-carnivorous-pitcher-plants-of-the-genus-nepenthes
#11
Michaela Saganová, Boris Bokor, Tibor Stolárik, Andrej Pavlovič
Nepenthes regulates enzyme activities by sensing stimuli from the insect prey. Protein is the best inductor mimicking the presence of an insect prey. Carnivorous plants of the genus Nepenthes have evolved passive pitcher traps for prey capture. In this study, we investigated the ability of chemical signals from a prey (chitin, protein, and ammonium) to induce transcription and synthesis of digestive enzymes in Nepenthes × Mixta. We used real-time PCR and specific antibodies generated against the aspartic proteases nepenthesins, and type III and type IV chitinases to investigate the induction of digestive enzyme synthesis in response to different chemical stimuli from the prey...
May 16, 2018: Planta
https://www.readbyqxmd.com/read/29764367/papain-like-and-legumain-like-proteases-in-rice-genome-wide-identification-comprehensive-gene-feature-characterization-and-expression-analysis
#12
Wei Wang, Xue-Mei Zhou, Han-Xian Xiong, Wan-Ying Mao, Peng Zhao, Meng-Xiang Sun
BACKGROUND: Papain-like and legumain-like proteases are proteolytic enzymes which play key roles in plant development, senescence and defense. The activities of proteases in both families could be inhibited by a group of small proteins called cystatin. Cystatin family genes have been well characterized both in tobacco and rice, suggesting their potential roles in seed development. However, their potential targets, papain-like and legumain-like proteases, have not been well characterized in plants, especially in rice, a model plant for cereal biology...
May 15, 2018: BMC Plant Biology
https://www.readbyqxmd.com/read/29763934/inactivation-of-usp14-perturbs-ubiquitin-homeostasis-and-delays-the-cell-cycle-in-mouse-embryonic-fibroblasts-and-in-fruit-fly-drosophila
#13
Jung Hoon Lee, Seoyoung Park, Yejin Yun, Won Hoon Choi, Min-Ji Kang, Min Jae Lee
BACKGROUND/AIMS: The 26S proteasome is the key proteolytic complex for recognition and degradation of polyubiquitinated target substrates in eukaryotes. Among numerous proteasome-associated proteins, a deubiquitinating enzyme (DUB) USP14 has been identified as an endogenous inhibitor of the proteasome. Here, we explored the complex regulatory functions of USP14 that involve ubiquitin (Ub) homeostasis and substrate degradation in flies and mammals. METHODS: USP14-null primary and immortalized mouse embryonic fibroblasts (MEFs) and USP14 knocked-down Drosophila were analyzed in this study...
May 9, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29758537/biopolymers-conjugated-with-magnetite-as-support-materials-for-trypsin-immobilization-and-protein-digestion
#14
Jakub Zdarta, Katarzyna Antecka, Artur Jędrzak, Karol Synoradzki, Magdalena Łuczak, Teofil Jesionowski
In the presented study synthesized magnetic nanoparticles were used as an inorganic precursor for the preparation of novel magnetite-lignin and magnetite-chitin hybrid supports for enzyme immobilization. Effective synthesis of the hybrids was confirmed by Fourier transform infrared spectroscopy and powder X-ray diffraction analysis. The materials exhibited good thermal stability and surface areas of 4.3 and 5.6 m2 /g respectively. The magnetite-lignin + trypsin and magnetite-chitin + trypsin systems were found to have good storage stability and reusability...
May 8, 2018: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29755650/preclinical-comparison-of-proteasome-and-ubiquitin-e1-enzyme-inhibitors-in-cutaneous-squamous-cell-carcinoma-the-identification-of-mechanisms-of-differential-sensitivity
#15
Angela McHugh, Kenneth Fernandes, Andrew P South, Jemima E Mellerio, Julio C Salas-Alanís, Charlotte M Proby, Irene M Leigh, Mark K Saville
Proteasome inhibitors have distinct properties and the biochemical consequences of suppressing ubiquitin E1 enzymes and the proteasome differ. We compared the effects of the proteasome inhibitors bortezomib, ixazomib and carfilzomib and the ubiquitin E1 enzyme inhibitor MLN7243/TAK-243 on cell viability and cell death in normal keratinocytes and cutaneous squamous cell carcinoma (cSCC) cell lines. The effects of both a pulse of treatment and more extended incubation were investigated. This is relevant to directly-delivered therapy (topical treatment/intratumoral injection) where the time of exposure can be controlled and a short exposure may better reflect systemically-delivered inhibitor pharmacokinetics...
April 17, 2018: Oncotarget
https://www.readbyqxmd.com/read/29751895/inhibitory-effects-of-chitosan-based-coatings-on-endogenous-enzyme-activities-proteolytic-degradation-and-texture-softening-of-grass-carp-ctenopharyngodon-idellus-fillets-stored-at-4-%C3%A2-c
#16
Dawei Yu, Joe M Regenstein, Jinhong Zang, Wenshui Xia, Yanshun Xu, Qixing Jiang, Fang Yang
The study evaluated the inhibitory effects of chitosan-based coatings on endogenous enzyme activities, proteolytic degradation and texture softening of grass carp (Ctenopharyngodon idellus) fillets stored at 4 °C for 15 days. In general, the inhibitory effects of chitosan coating were worse than that of chitosan composite coatings, which significantly inhibited cathepsin B, B + L and calpain activities of fillets, with the most reductions of 58%, 65% and 41%, respectively when compared to the corresponding values of control fillets...
October 1, 2018: Food Chemistry
https://www.readbyqxmd.com/read/29745033/role-of-teriparatide-in-glucocorticoid-induced-osteoporosis-through-regulating-cellular-reactive-oxygen-species
#17
Tao Wang, Chao Han, Peng Tian, Peng-Fei Li, Xin-Long Ma
OBJECTIVE: To determine the signaling pathways mediated by teriparatide in MLO-Y4 cell lines based on the evaluation of reactive oxygen species (ROS) through AKT pathways, which regulate apoptosis of bone cells. METHODS: We performed the DCFH-DA assay to investigate the role of ROS in MLO-Y4 cells caused by dexamethasone (Dex). Four groups were included: Dex group, Dex+NAC, Dex+ teriparatide group and control group (without any dispose). Real-time reverse transcriptase polymerase chain reaction was used to test the SOD2 and Cat mRNA expression...
May 10, 2018: Orthopaedic Surgery
https://www.readbyqxmd.com/read/29740840/antimicrobial-activity-of-an-aspartic-protease-from-salpichroa-origanifolia-fruits
#18
María Eugenia Díaz, Gabriela F Rocha, Francisco Kise, Adriana M Rosso, María Gabriela Guevara, Mónica G Parisi
Plant proteases play a fundamental rol in several processes like growth, development and in response to biotic and abiotic stress. In particular, aspartic proteases are expressed in different plant organs and have antimicrobial activity. Previously, we purified an aspartic protease from Salpichroa origanifolia fruits called salpichroin. The aim of this work was to determine the cytotoxic activity of this enzyme on selected plant and human pathogens. For this purpose, the growth of the selected pathogens was analysed after exposure to different concentrations of salpichroin...
May 8, 2018: Letters in Applied Microbiology
https://www.readbyqxmd.com/read/29735645/matrix-metalloproteinase-inhibitors-in-cancer-therapy-turning-past-failures-into-future-successes
#19
REVIEW
Arthur Winer, Sylvia Adams, Paolo Mignatti
The matrix metalloproteinases (MMP) are a family of proteolytic enzymes that degrade multiple components of the extracellular matrix. A large body of experimental and clinical evidence has implicated MMPs in tumor invasion, neoangiogenesis, and metastasis, and therefore they represent ideal pharmacologic targets for cancer therapy. From the 1990s to early 2000s, synthetic inhibitors of MMPs (MMPI) were studied in various cancer types. Unexpectedly, despite strongly promising preclinical data, all trials were unsuccessful in reducing tumor burden or improving overall survival; in addition, MMPIs had unforeseen, severe side effects...
May 7, 2018: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/29729742/inkjet-printing-based-%C3%AE-secretase-fluorescence-resonance-energy-transfer-fret-assay-for-screening-of-potential-%C3%AE-secretase-inhibitors-of-alzheimer-s-disease
#20
Jungmi Lee, Annie Agnes Suganya Samson, Joon Myong Song
Amyloid-β (Aβ) is generated by proteolytic processing of amyloid precursor protein (APP) by beta-secretase (BACE-1) and gamma-secretase. Amyloid-β is responsible for the formation of senile plaques in Alzheimer's disease (AD). Consequently, inhibition of β-secretase (BACE-1), a rate-limiting enzyme in the production of Aβ, constitutes an attractive therapeutic approach to the treatment of AD. This paper reports an inkjet printing-based fluorescence assay for high throughput screening of β-secretase inhibitors achieved by employing a BACE-1 FRET substrate (Rh-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys-Quencher)...
August 31, 2018: Analytica Chimica Acta
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