keyword
MENU ▼
Read by QxMD icon Read
search

Protein expression and purification

keyword
https://www.readbyqxmd.com/read/28918196/expression-and-purification-of-human-and-saccharomyces-cerevisiae-equilibrative-nucleoside-transporters
#1
Rebba C Boswell-Casteel, Jennifer M Johnson, Zygy Roe-Žurž, Kelli D Duggan, Hannah Schmitz, Franklin A Hays
Nucleosides play an essential role in the physiology of eukaryotes by acting as metabolic precursors in de novo nucleic acid synthesis and energy metabolism. Nucleosides also act as ligands for purinergic receptors. Equilibrative nucleoside transporters (ENTs) are polytopic integral membrane proteins that aid in regulating plasmalemmal flux of purine and pyrimidine nucleosides and nucleobases. ENTs exhibit broad substrate selectivity across different isoforms and utilize diverse mechanisms to drive substrate flux across membranes...
September 13, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28911643/cytotoxic-and-antioxidant-capacity-of-camel-milk-peptides-effects-of-isolated-peptide-on-superoxide-dismutase-and-catalase-gene-expression
#2
Masoud Homayouni-Tabrizi, Ahmad Asoodeh, Mozhgan Soltani
Peptides from natural sources such as milk are shown to have a wide spectrum of biological activities. In this study, three peptides with antioxidant capacity were identified from camel milk protein hydrolysate. Pepsin and pancreatin were used for hydrolysis of milk proteins. Ultrafiltration and reverse-phase high-performance liquid chromatography were used for the concentration and purification of the hydrolysate, respectively. Sequences of the three peptides, which were determined by matrix-assisted laser desorption/ionization time-of-flight spectrophotometry, were LEEQQQTEDEQQDQL [molecular weight (MW): 1860...
July 2017: Journal of Food and Drug Analysis
https://www.readbyqxmd.com/read/28911033/formulation-of-chitosan-with-the-polyepitope-hiv-1-protein-candidate-vaccine-efficiently-boosts-cellular-immune-responses-in-mice
#3
Ehsan Ollah Jazaeri, Atiyeh Mahdavi, Asghar Abdoli
Human immunodeficiency virus-1 (HIV-1) continues to be a major global public health issue and priority. Despite the variety of antiretroviral therapies, it seems that an effective vaccine against HIV-1 is still very necessary. An ideal HIV-1 vaccine should be able to elicit both humoral and cellular immunities. In this respect, polyepitope vaccines, incorporated from several conserved regions of HIV-1 proteins, have received much attention recently. Herein, the immunogenicity of the HIV-1 polyepitope protein-based candidate vaccines was evaluated in BALB/c mice...
November 30, 2017: Pathogens and Disease
https://www.readbyqxmd.com/read/28910620/ctag-paperclip-reveals-alternative-polyadenylation-promotes-cell-type-specific-protein-diversity-and-shifts-araf-isoforms-with-microglia-activation
#4
Hun-Way Hwang, Yuhki Saito, Christopher Y Park, Nathalie E Blachère, Yoko Tajima, John J Fak, Ilana Zucker-Scharff, Robert B Darnell
Alternative polyadenylation (APA) is increasingly recognized to regulate gene expression across different cell types, but obtaining APA maps from individual cell types typically requires prior purification, a stressful procedure that can itself alter cellular states. Here, we describe a new platform, cTag-PAPERCLIP, that generates APA profiles from single cell populations in intact tissues; cTag-PAPERCLIP requires no tissue dissociation and preserves transcripts in native states. Applying cTag-PAPERCLIP to profile four major cell types in the mouse brain revealed common APA preferences between excitatory and inhibitory neurons distinct from astrocytes and microglia, regulated in part by neuron-specific RNA-binding proteins NOVA2 and PTBP2...
September 13, 2017: Neuron
https://www.readbyqxmd.com/read/28904190/exploring-the-human-nipah-virus-protein-protein-interactions
#5
Luis Martinez-Gil, Natalia M Vera-Velasco, Ismael Mingarro
Nipah virus is an emerging, highly pathogenic, zoonotic virus of the paramyxoviridiae family. Human transmission occurs by close contact with infected animals, the consumption of contaminated food, or, occasionally, via other infected individuals. Currently, we lack therapeutic or prophylactic treatments for Nipah virus. To develop these agents we must now improve our understanding of the host-virus interactions that underpin a productive infection. This aim led us to perform the present work, in which we identified (101) human-Nipah virus protein-protein interactions (PPIs), most of which (88) are novel...
September 13, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28902877/purification-and-characterization-of-a-highly-specific-polyclonal-antibody-against-human-extracellular-signal-regulated-kinase-8-and-its-detection-in-lung-cancer
#6
Na-Li Cai, Andy T Y Lau, Fei-Yuan Yu, Dan-Dan Wu, Li-Juan Dai, Hai-Ying Mo, Chang-Min Lin, Yan-Ming Xu
Extracellular signal-regulated kinase 8 (ERK8), proposed as a novel potential therapeutic target for cancer, has been implicated in cell transformation, apoptosis, the protection of genomic integrity, and autophagy. To facilitate ERK8 research, a highly specific anti-ERK8 antibody is needed. In this article, we use the Immune Epitope Database and Analysis Resource online tool to predict B-cell epitopes of human ERK8 protein, and choose a 28 aa-peptide sequence to generate the GST-ERK8(28aa) fusion protein as the antigen for developing polyclonal antibody against ERK8...
2017: PloS One
https://www.readbyqxmd.com/read/28902865/optimization-of-the-expression-purification-and-polymerase-activity-reaction-conditions-of-recombinant-human-primpol
#7
Elizaveta O Boldinova, Gorazd Stojkovič, Rafil Khairullin, Sjoerd Wanrooij, Alena V Makarova
Human PrimPol is a DNA primase/polymerase involved in DNA damage tolerance and prevents nuclear genome instability. PrimPol is also localized to the mitochondria, but its precise function in mitochondrial DNA maintenance has remained elusive. PrimPol works both as a translesion (TLS) polymerase and as the primase that restarts DNA replication after a lesion. However, the observed biochemical activities of PrimPol vary considerably between studies as a result of different reaction conditions used. To reveal the effects of reaction composition on PrimPol DNA polymerase activity, we tested the polymerase activity in the presence of various buffer agents, salt concentrations, pH values and metal cofactors...
2017: PloS One
https://www.readbyqxmd.com/read/28901644/the-truncated-nlr-protein-tir-nbs13-is-a-mos6-importin-%C3%AE-3-interaction-partner-and-required-for-plant-immunity
#8
Charlotte Roth, Daniel Lüdke, Melanie Klenke, Annalena Quathamer, Oliver Valerius, Gerhard H Braus, Marcel Wiermer
Importin-α proteins mediate the translocation of nuclear localization signal (NLS)-containing proteins from the cytoplasm into the nucleus through nuclear pore complexes (NPCs). Genetically, Arabidopsis IMPORTIN-α3/MOS6 (MODIFIER OF SNC1, 6) is required for basal plant immunity and constitutive disease resistance activated in the auto-immune mutant snc1 (suppressor of npr1-1, constitutive 1), suggesting that MOS6 plays a role in nuclear import of proteins involved in plant defense signaling. Here, we sought to identify and characterize defense-regulatory cargo proteins and interaction partners of MOS6...
September 13, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28900375/expression-and-purification-of-untagged-glnk-proteins-from-actinobacteria
#9
Edileusa C M Gerhardt, Vivian R Moure, Andrey W Souza, Fabio O Pedrosa, Emanuel M Souza, Lautaro Diacovich, Hugo Gramajo, Luciano F Huergo
The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been well documented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficient purification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor...
2017: EXCLI journal
https://www.readbyqxmd.com/read/28895884/interaction-of-recombinant-gallus-gallus-sept5-and-brain-proteins-of-h5n1-avian-influenza-virus-infected-chickens
#10
Jasmine Elanie Khairat, Vinod Balasubramaniam, Iekhsan Othman, Abdul Rahman Omar, Sharifah Syed Hassan
Septin forms a conserved family of cytoskeletal guanosine triphosphate (GTP) binding proteins that have diverse roles in protein scaffolding, vesicle trafficking, and cytokinesis. The involvement of septins in infectious viral disease pathogenesis has been demonstrated by the upregulation of SEPT5 protein and its mRNA in brain tissues of H5N1-infected chickens, thus, providing evidence for the potential importance of this protein in the pathogenesis of neurovirulence caused by the avian influenza virus. In this study, cloning, expression, and purification of Gallus gallus SEPT5 protein was performed in Escherichia coli...
September 12, 2017: Proteomes
https://www.readbyqxmd.com/read/28895364/-development-of-tumor-targeting-phbhhx-nanoparticles-by-phap-mediated-immobilization-of-egfr-targeting-peptide
#11
Fan Fan, Jiangang Ma, Xiaoyun Lu, Teng Niu, Zhimin Wu
PHA granule binding protein phasin (PhaP) has a high affinity for hydrophobic materials and can bind to hydrophobic polymers via strong hydrophobic interaction. In this study, an EGFR-targeting peptide (ETP) was fused with PhaP and the fusion protein ETP-PhaP was produced in recombinant Escherichia coli BL21 (DE3) (pPI-ETP-P) and then purified by Ni affinity purification. The tumor targeting PHBHHx nanoparticles were developed based on PhaP mediated ETP immobilization and the cellular uptake of the ETP-PhaP modified PHBHHx NPs and none modified PHBHHx NPs by cervical cancer cell lines SiHa (EGFR over expressed) and CaSKi (EGFR low expressed) were analyzed...
June 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28894144/molecular-details-of-secretory-phospholipase-a2-from-flax-linum-usitatissimum-l-provide-insight-into-its-structure-and-function
#12
Payal Gupta, Prasanta K Dash
Secretory phospholipase A2 (sPLA2) are low molecular weight proteins (12-18 kDa) involved in a suite of plant cellular processes imparting growth and development. With myriad roles in physiological and biochemical processes in plants, detailed analysis of sPLA2 in flax/linseed is meagre. The present work, first in flax, embodies cloning, expression, purification and molecular characterisation of two distinct sPLA2s (I and II) from flax. PLA2 activity of the cloned sPLA2s were biochemically assayed authenticating them as bona fide phospholipase A2...
September 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28893289/recombinant-expression-of-beak-and-feather-disease-virus-capsid-protein-and-assembly-of-virus-like-particles-in-nicotiana-benthamiana
#13
Guy L Regnard, Edward P Rybicki, Inga I Hitzeroth
BACKGROUND: Beak and feather disease virus (BFDV) is an important disease causing agent affecting psittacines. BFDV is highly infectious and can present as acute, chronic or subclinical disease. The virus causes immunodeficiency and is often associated with secondary infections. No commercial vaccine is available and yields of recombinant BFDV capsid protein (CP) expressed in insect cells and bacteria are yet to be seen as commercially viable, although both systems produced BFDV CP that could successfully assemble into virus-like particles (VLPs)...
September 11, 2017: Virology Journal
https://www.readbyqxmd.com/read/28890895/isolation-of-fibroblast-activation-protein-specific-cancer-associated-fibroblasts
#14
Yingying Huang, Sufang Zhou, Yong Huang, Duo Zheng, Qiqi Mao, Jian He, Yiwei Wang, Dabing Xue, Xiaoling Lu, Nuo Yang, Yongxiang Zhao
The current study is to develop a gentle and efficient method for purification of fibroblast-activation protein positive (FAP(+)) cancer-associated fibroblasts (CAFs) from tumor tissues. Fresh tissues were isolated from BALB/c-Nude mice bearing human liver cancer cell line (HepG2), fully minced and separated into three parts, and digested with trypsin digestion and then treated with collagenase type IV once, twice, or thrice, respectively. Finally, the cells were purified by using FAP magnetic beads. The isolated CAFs were grown in culture medium and detected for the surface expression of fibroblast-activation protein (FAP)...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28888757/single-step-purification-of-recombinant-gaussia-luciferase-from-serum-containing-culture-medium-of-mammalian-cells
#15
Satoshi Inouye
A dihydrofolate reductase-deficient Chinese hamster ovary (CHO-K1/dhfr(-)) cell line stably expressing Gaussia luciferase with a histidine-tag sequence at the carboxyl terminus (GLase-His) was established. Recombinant GLase-His was purified from serum-containing culture medium by single-step Ni-chelate column chromatography in the presence of 2 M NaCl and 0.01% Tween 20. The protein yield of GLase-His with over 95% purity was 0.5 mg from 0.9 L of the cultured medium. The enzymatic properties of purified GLase-His were characterized...
September 6, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28886312/differences-in-expression-of-serum-protein-in-patients-with-psoriasis-vulgaris-and-blood-heat-syndrome-and-healthy-volunteers
#16
P Hao, C Yang, X Yan, T Li, J Zhang, Q Yu, X Zhang, Y Zhang
To explore the mechanism of psoriasis vulgaris (PV), serum protein expression profiles between PV patients with blood-heat syndrome and healthy volunteers were detected by isobaric tags for relative and absolute quantitation (iTRAQ). First, sera from 15 PV patients with blood-heat syndrome and 10 healthy volunteers were collected; then, serum proteins were separated and hydrolyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a specific iTRAQ marker enzyme respectively after further purification and protein abundance treatment...
August 30, 2017: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/28883622/socs2-binds-to-and-regulates-epha2-through-multiple-mechanisms
#17
Carissa Pilling, Jonathan A Cooper
Suppressors of cytokine signaling (SOCS) proteins inhibit signaling by serving as substrate receptors for the Cullin5-RING E3 ubiquitin ligase (CRL5) and through a variety of CRL5-independent mechanisms. CRL5, SOCS2 and SOCS6 are implicated in suppressing transformation of epithelial cells. We identified cell proteins that interact with SOCS2 and SOCS6 using two parallel proteomics techniques: BioID and Flag affinity purification mass spectrometry. The receptor tyrosine kinase ephrin type-A receptor 2 (EphA2) was identified as a SOCS2-interacting protein...
September 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28882309/expression-and-isolation-of-recombinant-tau
#18
Rebecca L Best, Peter J Chung, Sarah J Benbow, April Savage, Nichole E LaPointe, Cyrus R Safinya, Stuart C Feinstein
In this chapter, we describe methods for the purification of both untagged and polyhistidine-tagged tau protein. These protocols utilize a bacterial expression system to produce the tau isoform of interest, followed by heat treatment and column chromatography to separate tau from impurities. These techniques yield a biochemically pure protein with which to pursue any number of questions regarding the mechanisms of tau action.
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28882203/nitrogenase-assembly-strategies-and-procedures
#19
Nathaniel S Sickerman, Yilin Hu, Markus W Ribbe
Nitrogenase is a metalloenzyme system that plays a critical role in biological nitrogen fixation, and the study of how its metallocenters are assembled into functional entities to facilitate the catalytic reduction of dinitrogen to ammonia is an active area of interest. The diazotroph Azotobacter vinelandii is especially amenable to culturing and genetic manipulation, and this organism has provided the basis for many insights into the assembly of nitrogenase proteins and their respective metallocofactors. This chapter will cover the basic procedures necessary for growing A...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28880952/large-scale-expression-and-purification-of-secreted-mouse-hephaestin
#20
Chandrika N Deshpande, Vicky Xin, Yan Lu, Tom Savage, Gregory J Anderson, Mika Jormakka
Hephaestin is a large membrane-anchored multicopper ferroxidase involved in mammalian iron metabolism. Newly absorbed dietary iron is exported across the enterocyte basolateral membrane by the ferrous iron transporter ferroportin, but hephaestin increases the efficiency of this process by oxidizing the transported iron to its ferric form and promoting its release from ferroportin. Deletion or mutation of the hephaestin gene leads to systemic anemia with iron accumulation in the intestinal epithelium. The crystal structure of human ceruloplasmin, another multicopper ferroxidase with 50% sequence identity to hephaestin, has provided a framework for comparative analysis and modelling...
2017: PloS One
keyword
keyword
58882
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"