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membrane protein purification

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https://www.readbyqxmd.com/read/27890709/role-of-major-and-brain-specific-sgce-isoforms-in-the-pathogenesis-of-myoclonus-dystonia-syndrome
#1
Jianfeng Xiao, Satya R Vemula, Yi Xue, Mohammad M Khan, Francesca A Carlisle, Adrian J Waite, Derek J Blake, Ioannis Dragatsis, Yu Zhao, Mark S LeDoux
Loss-of-function mutations in SGCE, which encodes ε-sarcoglycan (ε-SG), cause myoclonus-dystonia syndrome (OMIM159900, DYT11). A "major" ε-SG protein derived from CCDS5637.1 (NM_003919.2) and a "brain-specific" protein, that includes sequence derived from alternative exon 11b (CCDS47642.1, NM_001099400.1), are reportedly localized in post- and pre-synaptic membrane fractions, respectively. Moreover, deficiency of the "brain-specific" isoform and other isoforms derived from exon 11b may be central to the pathogenesis of DYT11...
November 24, 2016: Neurobiology of Disease
https://www.readbyqxmd.com/read/27890399/a-partially-purified-outer-membrane-protein-virb9-1-for-low-cost-nanovaccines-against-anaplasma-marginale
#2
Liang Zhao, Antonino S Cavallaro, David Wibowo, Bing Zhang, Jun Zhang, Neena Mitter, Chengzhong Yu, Chun-Xia Zhao, Anton P J Middelberg
Anaplasma marginale is a devastating tick-borne pathogen causing anaplasmosis in cattle and results in significant economic loss to the cattle industry worldwide. Currently, there is no widely accepted vaccine against A. marginale. New generation subunit vaccines against A. marginale, which are much safer, more efficient and cost-effective, are in great need. The A. marginale outer membrane protein VirB9-1 is a promising antigen for vaccination. We previously have shown that soluble recombinant VirB9-1 protein can be expressed and purified from Escherichia coli and induce a high level of humoral and cellular immunity in mice...
November 24, 2016: Vaccine
https://www.readbyqxmd.com/read/27873257/bacterial-histidine-kinases-overexpression-purification-and-inhibitor-screen
#3
Mike Gajdiss, Michael Türck, Gabriele Bierbaum
Bacterial histidine kinases are promising targets for new antimicrobial agents. In antibacterial therapy such agents could inhibit bacterial growth by targeting essential two-component regulatory systems or resensitize bacteria to known antibiotics by blocking stress responses like the cell wall stress response. However, (1) activity assays using the truncated phosphorylation domains have been shown to produce artifacts and (2) the purification of the full-length histidine kinases is complicated. Here, we describe a standard protocol for the recombinant expression and purification of functional full-length histidine kinases and other membrane proteins from gram-positive bacteria that do not harbor more than two trans-membrane domains using an Escherichia coli host...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27862975/purification-of-soluble-hla-class-i-complexes-from-human-serum-or-plasma-deliver-high-quality-immuno-peptidomes-required-for-biomarker-discovery
#4
Danilo Ritz, Andreas Gloger, Dario Neri, Tim Fugmann
Soluble human leukocyte antigen class I (sHLA)-peptide complexes have been suggested to play a role in the modulation of immune responses and in immune evasion of cancer cells. The set of peptides eluted from sHLA molecules could serve as biomarker for the monitoring of patients with cancer or other conditions. Here, we describe an improved sHLA peptidomics methodology resulting in the identification of 1816 to 2761 unique peptide sequences from triplicate analyses of serum or plasma taken from three healthy donors...
November 12, 2016: Proteomics
https://www.readbyqxmd.com/read/27859689/protection-against-genital-tract-chlamydia-trachomatis-infection-following-intranasal-immunization-with-a-novel-recombinant-momp-vs2-4-antigen
#5
Ronza Hadad, Ellen Marks, Irina Kalbina, Karin Schön, Magnus Unemo, Nils Lycke, Åke Strid, Sören Andersson
The asymptomatic nature of most Chlamydia trachomatis infections and the lack of appropriate effects by current prevention and management call for vaccine development. We evaluated a recombinant subunit vaccine candidate based on the major outer membrane protein variable segments 2 and 4 (MOMP VS2/4). To achieve maximal immunogenicity and ease of production and purification, MOMP VS2/4 was constructed by using highly immunogenic sequences of MOMP only, thereby minimizing the presence of hydrophobic regions, and spacing the immunogenic epitopes with a flexible amino acid sequence...
December 2016: APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica
https://www.readbyqxmd.com/read/27858178/proteome-analysis-of-hemofilter-adsorbates-to-identify-novel-substances-of-sepsis-a-pilot-study
#6
Tomoaki Hashida, Taka-Aki Nakada, Mamoru Satoh, Keisuke Tomita, Rui Kawaguchi, Fumio Nomura, Shigeto Oda
Blood purification therapy using hemofilters with high adsorbing capabilities has been reported to remove excessive humoral mediators from the blood of patients with sepsis. However, there are insufficient studies of the adsorbates bound to hemofilter membranes. We hypothesized that these adsorbates in acute kidney injury (AKI) patients with sepsis were different from those in patients without sepsis and that proteome analysis of the adsorbates would identify novel substances of sepsis. This study included 20 patients who had AKI upon admission to intensive care units (ICUs) and who received continuous renal replacement therapy using polymethyl methacrylate hemofilters...
November 17, 2016: Journal of Artificial Organs: the Official Journal of the Japanese Society for Artificial Organs
https://www.readbyqxmd.com/read/27854364/deep-interactome-profiling-of-membrane-proteins-by-co-interacting-protein-identification-technology
#7
Sandra Pankow, Casimir Bamberger, Diego Calzolari, Andreas Bamberger, John R Yates
Affinity purification coupled to mass spectrometry (AP-MS) is the method of choice for analyzing protein-protein interactions, but common protocols frequently recover only the most stable interactions and tend to result in low bait yield for membrane proteins. Here, we present a novel, deep interactome sequencing approach called CoPIT (co-interacting protein identification technology), which allows comprehensive identification and analysis of membrane protein interactomes and their dynamics. CoPIT integrates experimental and computational methods for a coimmunoprecipitation (Co-IP)-based workflow from sample preparation for mass spectrometric analysis to visualization of protein-protein interaction networks...
December 2016: Nature Protocols
https://www.readbyqxmd.com/read/27847172/novel-systematic-detergent-screening-method-for-membrane-proteins-solubilization
#8
Elodie Desuzinges Mandon, Morgane Agez, Rebecca Pellegrin, Sébastien Igonet, Anass Jawhari
Membrane proteins play crucial role in many cellular processes including cell adhesion, cell-cell communication, signal transduction and transport. To better understand the molecular basis of such central biological machines and in order to specifically study their biological and medical role, it is necessary to extract them from their membrane environment. To do so, it is challenging to find the best solubilization condition. Here we describe, a systematic screening method called BMSS (Biotinylated Membranes Solubilization & Separation) that allow screening 96 conditions at once...
November 12, 2016: Analytical Biochemistry
https://www.readbyqxmd.com/read/27845784/antibody-purification-via-affinity-membrane-chromatography-method-utilizing-nucleotide-binding-site-targeting-with-a-small-molecule
#9
Nur Mustafaoglu, Tanyel Kiziltepe, Basar Bilgicer
Here, we present an affinity membrane chromatography technique for purification of monoclonal and polyclonal antibodies from cell culture media of hybridomas and ascites fluids. The m-NBST method utilizes the nucleotide-binding site (NBS) that is located on the Fab variable domain of immunoglobulins to enable capturing of antibody molecules on a membrane affinity column via a small molecule, tryptamine, which has a moderate binding affinity to the NBS. Regenerated cellulose membrane was selected as a matrix due to multiple advantages over traditionally used resin-based affinity systems...
November 15, 2016: Analyst
https://www.readbyqxmd.com/read/27837483/characterization-of-type-three-secretion-system-translocator-interactions-with-phospholipid-membranes
#10
Philip R Adam, Michael L Barta, Nicholas E Dickenson
In vitro characterization of type III secretion system (T3SS) translocator proteins has proven challenging due to complex purification schemes and their hydrophobic nature that often requires detergents to provide protein solubility and stability. Here, we provide experimental details for several techniques that overcome these hurdles, allowing for the direct characterization of the Shigella translocator protein IpaB with respect to phospholipid membrane interaction. The techniques specifically discussed in this chapter include membrane interaction/liposome flotation, liposome sensitive fluorescence quenching, and protein-mediated liposome disruption assays...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27825980/expression-and-purification-of-native-and-functional-influenza-a-virus-matrix-2-proton-selective-ion-channel
#11
Elodie Desuzinges Mandon, Aurélien Traversier, Anne Champagne, Lorraine Benier, Stéphane Audebert, Sébastien Balme, Emmanuel Dejean, Manuel Rosa Calatrava, Anass Jawhari
Influenza A virus displays one of the highest infection rates of all human viruses and therefore represents a severe human health threat associated with an important economical challenge. Influenza matrix protein 2 (M2) is a membrane protein of the viral envelope that forms a proton selective ion channel. Here we report the expression and native isolation of full length active M2 without mutations or fusions. The ability of the influenza virus to efficiently infect MDCK cells was used to express native M2 protein...
November 5, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/27815048/electrophysiological-activity-of-a-neurotoxic-fraction-from-the-venom-of-box-jellyfish-carybdea-marsupialis
#12
Fernando Lazcano-Pérez, Rogelio O Arellano, Edith Garay, Roberto Arreguín-Espinosa, Judith Sánchez-Rodríguez
Carybdea marsupialis is a widely distributed box jellyfish found in the Mediterranean and in the tropical waters of the Caribbean Sea. Its venom is a complex mixture of biologically active compounds that are used to catch prey. In order to evaluate the activity of the neurotoxins in the venom, bioassays were carried out using the marine crab Ocypode quadrata. The proteins with neurotoxic effect were partially purified using low-pressure liquid chromatography techniques. Gel filtration (Sephadex G-50M) was used as the first step and the active fraction in crabs was passed through a QAE Sephadex A-25 column...
November 1, 2016: Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP
https://www.readbyqxmd.com/read/27813588/purification-biochemical-characterization-and-antimicrobial-activity-of-a-new-lipid-transfer-protein-from-coffea-canephora-seeds
#13
G C V Bard, U Zottich, T A M Souza, S F F Ribeiro, G B Dias, S Pireda, M Da Cunha, R Rodrigues, L S Pereira, O L T Machado, A O Carvalho, V M Gomes
Coffee, an agronomical crop of great economic importance, is also among the most commonly traded commodities in worldwide markets. Antimicrobial peptides, which play a role in plant defense, have been identified and isolated particularly from seeds. We isolated and immunolocalized Cc-LTP2, a new lipid transfer protein (LTP) from Coffea canephora seeds. We report its antimicrobial activity against various phytopathogenic fungi of economic importance, and against the bacterium Xanthomonas euvesicatoria. Peptides from C...
October 24, 2016: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/27807400/aqueous-two-phase-system-atps-an-overview-and-advances-in-its-applications
#14
REVIEW
Mujahid Iqbal, Yanfei Tao, Shuyu Xie, Yufei Zhu, Dongmei Chen, Xu Wang, Lingli Huang, Dapeng Peng, Adeel Sattar, Muhammad Abu Bakr Shabbir, Hafiz Iftikhar Hussain, Saeed Ahmed, Zonghui Yuan
Aqueous two-phase system (ATPS) is a liquid-liquid fractionation technique and has gained an interest because of great potential for the extraction, separation, purification and enrichment of proteins, membranes, viruses, enzymes, nucleic acids and other biomolecules both in industry and academia. Although, the partition behavior involved in the method is complex and difficult to predict. Current research shows that it has also been successfully used in the detection of veterinary drug residues in food, separation of precious metals, sewage treatment and a variety of other purposes...
2016: Biological Procedures Online
https://www.readbyqxmd.com/read/27801621/map-tag-a-novel-tagging-system-for-protein-purification-and-detection
#15
Yuki Fujii, Mika K Kaneko, Yukinari Kato
Protein purification is an essential procedure in fields such as biochemistry, molecular biology, and biophysics. Acquiring target proteins with high quality and purity is still difficult, although several tag systems have been established for protein purification. Affinity tag systems are excellent because they possess high affinity and specificity for acquiring the target proteins. Nevertheless, further affinity tag systems are needed to compensate for several disadvantages of the presently available affinity tag systems...
November 1, 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/27799318/independent-activation-of-ion-conduction-pores-in-the-double-barreled-calcium-activated-chloride-channel-tmem16a
#16
Novandy K Lim, Andy K M Lam, Raimund Dutzler
The TMEM16 proteins constitute a family of membrane proteins with unusual functional breadth, including lipid scramblases and Cl(-) channels. Members of both these branches are activated by Ca(2+), acting from the intracellular side, and probably share a common architecture, which was defined in the recent structure of the lipid scramblase nhTMEM16. The structural features of subunits and the arrangement of Ca(2+)-binding sites in nhTMEM16 suggest that the dimeric protein harbors two locations for catalysis that are independent with respect to both activation and lipid conduction...
November 2016: Journal of General Physiology
https://www.readbyqxmd.com/read/27790201/changes-in-serological-immunology-measures-in-uk-and-kenyan-adults-post-controlled-human-malaria-infection
#17
Susanne H Hodgson, David Llewellyn, Sarah E Silk, Kathryn H Milne, Sean C Elias, Kazutoyo Miura, Gathoni Kamuyu, Elizabeth A Juma, Charles Magiri, Alfred Muia, Jing Jin, Alexandra J Spencer, Rhea J Longley, Thomas Mercier, Laurent Decosterd, Carole A Long, Faith H Osier, Stephen L Hoffman, Bernhards Ogutu, Adrian V S Hill, Kevin Marsh, Simon J Draper
Background: The timing of infection is closely determined in controlled human malaria infection (CHMI) studies, and as such they provide a unique opportunity to dissect changes in immunological responses before and after a single infection. The first Kenyan Challenge Study (KCS) (Pan African Clinical Trial Registry: PACTR20121100033272) was performed in 2013 with the aim of establishing the CHMI model in Kenya. This study used aseptic, cryopreserved, attenuated Plasmodium falciparum sporozoites administered by needle and syringe (PfSPZ Challenge) and was the first to evaluate parasite dynamics post-CHMI in individuals with varying degrees of prior exposure to malaria...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27787842/immunoaffinity-purification-of-the-glycosylated-extracellular-fragment-of-mouse-plexin-a2-produced-in-a-mammalian-expression-system
#18
Terukazu Nogi, Emiko Mihara, Norihisa Yasui, Junichi Takagi
Plexins are type I membrane proteins that function as receptors for semaphorins. All of the known plexins contain a large globular domain, termed the sema domain, in the N-terminal extracellular region, which interacts with semaphorins during signal transduction. Here, we describe procedures for protein production and purification that we utilized in the crystallographic study of the mouse Plexin A2 (mPlxnA2) extracellular fragment, including the sema domain. A mutant mammalian cell line, HEK293S GnTI(-), was used as an expression host for the production of a crystallizable-quality mPlxnA2 fragment, which contains several N-glycosylation sites and disulfide bonds...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27773917/high-cell-density-cultivation-of-nitrosomonas-europaea-in-a-membrane-bioreactor-for-performing-protein-purification-and-characterization-studies
#19
Mamoru Oshiki, Ryota Takagi, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki
No abstract text is available yet for this article.
October 21, 2016: Journal of General and Applied Microbiology
https://www.readbyqxmd.com/read/27773766/affinity-purification-of-bacterial-outer-membrane-vesicles-omvs-utilizing-a-his-tag-mutant
#20
Nathan J Alves, Kendrick B Turner, Kyle A DiVito, Michael A Daniele, Scott A Walper
To facilitate the rapid purification of bacterial outer membrane vesicles (OMVs), we developed two plasmid constructs that utilize a truncated, transmembrane protein to present an exterior histidine repeat sequence. We chose OmpA, a highly abundant porin protein, as the protein scaffold and utilized the lac promoter to allow for inducible control of the epitope-presenting construct. OMVs containing mutant OmpA-His6 were purified directly from Escherichia coli culture media on an immobilized metal affinity chromatography (IMAC) Ni-NTA resin...
October 20, 2016: Research in Microbiology
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