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Keywords Extraction and purification of...

Extraction and purification of membrane proteins

https://read.qxmd.com/read/38419296/endophytic-streptomyces-sp-msare05-isolated-from-roots-of-peanut-plant-produces-a-novel-antimicrobial-compound
#1
JOURNAL ARTICLE
Md Majharul Islam, Shrabani Saha, Prithidipa Sahoo, Sukhendu Mandal
AIM: This study aimed to isolate, endophytic Streptomyces sp. MSARE05 isolated from root of a peanut (Arachis hypogaea) inhibits the growth of other bacteria. The research focused on characterizing the strain and the antimicrobial compound. METHODS AND RESULTS: The surface sterilized peanut roots were used to isolate the endophytic bacterium Streptomyces sp. MSARE05. A small-scale fermentation was done to get the antimicrobial compound SM05 produced in highest amount in ISP-2 medium (pH 7) for 7 days at 30°C in shaking (180 rpm) condition...
February 28, 2024: Journal of Applied Microbiology
https://read.qxmd.com/read/38408346/anodic-electrodepositing-bioinspired-cu-bdc-nh-2-graphene-oxide-membrane-for-efficient-uranium-extraction
#2
JOURNAL ARTICLE
Bing Yao, Zhou Fang, Yue Hu, Zhizhen Ye, Xinsheng Peng
The challenge of removing trace levels of heavy metal ions, particularly uranium, from wastewater is a critical concern in environmental management. Uranium, a key element in long-term nuclear power generation, often poses significant extraction difficulties in wastewater due to its low concentration, interference from other ions, and the complexity of aquatic ecosystems. This study introduces an anodic electrodeposited hierarchical porous 2D metal-organic framework (MOF) Cu-BDC-NH2 @graphene oxide (GO) membrane for effective uranium extraction by mimicking the function of the superb-uranyl-binding protein...
February 26, 2024: Langmuir: the ACS Journal of Surfaces and Colloids
https://read.qxmd.com/read/38405833/a-proteome-wide-quantitative-guide-for-nanoscale-spatially-resolved-extraction-of-membrane-proteins-into-native-nanodiscs
#3
Caroline Brown, Snehasish Ghosh, Rachel McAllister, Mukesh Kumar, Gerard Walker, Eric Sun, Talat Aman, Aniruddha Panda, Shailesh Kumar, Wenxue Li, Jeff Coleman, Yansheng Liu, James E Rothman, Moitrayee Bhatacharyya, Kallol Gupta
The intricate molecular environment of the native membrane profoundly influences every aspect of membrane protein (MP) biology. Despite this, the most prevalent method of studying MPs uses detergent- like molecules that disrupt and remove this vital local membrane context. This severely impedes our ability to quantitatively decipher the local molecular context and comprehend its regulatory role in the structure, function, and biogenesis of MPs. Using a library of membrane-active polymers we have developed a platform for the high-throughput analysis of the membrane proteome...
February 12, 2024: bioRxiv
https://read.qxmd.com/read/38396666/-brassica-oleracea-l-var-italica-aquaporin-reconstituted-proteoliposomes-as-nanosystems-for-resveratrol-encapsulation
#4
JOURNAL ARTICLE
Lucia Yepes-Molina, José A Teruel, Urban Johanson, Micaela Carvajal
Aquaporins (AQPs), membrane proteins responsible for facilitating water transport, found in plant membrane vesicles (MV), have been related to the functionality and stability of MV. We focused on AQPs obtained from broccoli, as they show potential for biotechnological applications. To gain further insight into the role of AQPs in MV, we describe the heterologous overexpression of two broccoli AQPs ( BoPIP1;2 and BoPIP2;2 ) in Pichia pastoris , resulting in their purification with high yield (0.14 and 0.99 mg per gram cells for BoPIP1;2 and BoPIP2;2)...
February 6, 2024: International Journal of Molecular Sciences
https://read.qxmd.com/read/38395124/purification-and-characterisation-of-the-platelet-activating-gpvi-fcr%C3%AE-complex-in-smalps
#5
JOURNAL ARTICLE
Xueqing Wang, Alexandre Slater, Sarah C Lee, Neale Harrison, Naomi L Pollock, Saskia E Bakker, Stefano Navarro, Bernhard Nieswandt, Tim R Dafforn, Ángel García, Steve P Watson, Michael G Tomlinson
The collagen/fibrin(ogen) receptor, glycoprotein VI (GPVI), is a platelet activating receptor and a promising anti-thrombotic drug target. However, while agonist-induced GPVI clustering on platelet membranes has been shown to be essential for its activation, it is unknown if GPVI dimerisation represents a unique conformation for ligand binding. Current GPVI structures all contain only the two immunoglobulin superfamily (IgSF) domains in the GPVI extracellular region, so lacking the mucin-like stalk, transmembrane, cytoplasmic tail of GPVI and its associated Fc receptor γ (FcRγ) homodimer signalling chain, and provide contradictory insights into the mechanisms of GPVI dimerisation...
February 21, 2024: Archives of Biochemistry and Biophysics
https://read.qxmd.com/read/38390965/enhanced-electrophoretic-depletion-of-sodium-dodecyl-sulfate-with-methanol-for-membrane-proteome-analysis-by-mass-spectrometry
#6
JOURNAL ARTICLE
Hammam H Said, Alan A Doucette
Membrane proteins are underrepresented during proteome characterizations, primarily owing to their lower solubility. Sodium dodecyl sulfate (SDS) is favored to enhance protein solubility but interferes with downstream analysis by mass spectrometry. Here, we present an improved workflow for SDS depletion using transmembrane electrophoresis (TME) while retaining a higher recovery of membrane proteins. Though higher levels of organic solvent lower proteome solubility, we found that the inclusion of 40% methanol provided optimal solubility of membrane proteins, with 86% recovery relative to extraction with SDS...
February 2, 2024: Proteomes
https://read.qxmd.com/read/38347414/differential-glycoform-analysis-of-muc1-derived-from-biological-specimens-using-an-antibody-overlay-lectin-microarray
#7
JOURNAL ARTICLE
Atsushi Matsuda, Patcharaporn Boottanun, Sachiko Koizumi, Misugi Nagai, Atsushi Kuno
The association between altered glycosylation of MUC1 and various disease events has sparked significant interest. However, analytical technologies to investigate the disease-related glycoforms of endogenous MUC1 in blood and tissue specimens are limited. Therefore, we devised a reliable technique for differential analysis of endogenous MUC1 glycoforms based on an antibody-assisted lectin microarray. Its highly sensitive detection aids in analyzing soluble MUC1 from relatively small amounts of serum via a simple enrichment process...
2024: Methods in Molecular Biology
https://read.qxmd.com/read/38311972/hplc-purification-of-antioxidant-and-antibacterial-peptides-from-a-lichen-parmotrema-perlatum-huds-m-choisy-identification-by-lc-ms-ms-peptide-mass-fingerprinting
#8
JOURNAL ARTICLE
Dwarakanath P Radhakrishnan, Abinaya Kanakaraja, Nagasathiya Krishnan, Meenakumari Sakthivelu, Subash C B Gopinath, Raman Pachaiappan
Parmotrema perlatum, a lichen belonging to the family Parmeliaceae, is well known for its culinary benefits and aroma used as a condiment in Indian homes is also known as the "black stone flower" or "kalpasi" in India. This research intends to analyze the antioxidant power of the crude extracts using four pH-based buffers solubilized proteins/peptides and RP-HPLC fractions of P. perlatum obtained by purification. The proteins that were extracted from the four different buffers were examined using LC-MS/MS-based peptide mass fingerprinting...
February 4, 2024: Biotechnology and Applied Biochemistry
https://read.qxmd.com/read/38217646/extraction-and-purification-of-outer-membrane-vesicles-and-their-associated-rnas
#9
JOURNAL ARTICLE
Anaïs Blache, Wafa Achouak
Outer membrane vesicles (OMVs), produced by Gram negative-bacteria and sRNAs, are key players in cell-to-cell communication and interactions of bacteria with the environment. OMVs act as information carriers and encapsulate various molecules such as proteins, lipids, metabolites, and RNAs. OMVs and sRNAs play a broad range of functions from pathogenesis to stress resistance, to biofilm formation and both mediate interkingdom signaling. Various studies indicate that there is a mechanism of intercellular communication mediated by OMV-derived bacterial RNAs that is conserved among certain bacterial species...
2024: Methods in Molecular Biology
https://read.qxmd.com/read/38158037/purification-and-characterization-of-cdr1-the-drug-efflux-pump-conferring-azole-resistance-in-candida-species
#10
JOURNAL ARTICLE
Jorgaq Pata, Alexis Moreno, Benjamin Wiseman, Sandrine Magnard, Idriss Lehlali, Marie Dujardin, Atanu Banerjee, Martin Högbom, Ahcène Boumendjel, Vincent Chaptal, Rajendra Prasad, Pierre Falson
Candida albicans and C. glabrata express exporters of the ATP-binding cassette (ABC) superfamily and address them to their plasma membrane to expel azole antifungals, which cancels out their action and allows the yeast to become multidrug resistant (MDR). In a way to understand this mechanism of defense, we describe the purification and characterization of Cdr1, the membrane ABC exporter mainly responsible for such phenotype in both species. Cdr1 proteins were functionally expressed in the baker yeast, tagged at their C-terminal end with either a His-tag for the glabrata version, cgCdr1-His, or a green fluorescent protein (GFP) preceded by a proteolytic cleavage site for the albicans version, caCdr1-P-GFP...
December 27, 2023: Biochimie
https://read.qxmd.com/read/38066481/selective-extraction-of-recombinant-membrane-proteins-from-hansenula-polymorpha-by-pulsed-electric-field-and-lytic-enzyme-pretreatment
#11
JOURNAL ARTICLE
Valentina Ganeva, Andreas Kranz
BACKGROUND: In yeast, recombinant membrane proteins including viral scaffold proteins used for the formation of enveloped Virus-like particles (eVLPs) typically accumulate intracellularly. Their recovery is carried out by mechanical disruption of the cells, often in combination with detergent treatment. Cell permeabilization is an attractive alternative to mechanical lysis because it allows for milder and more selective recovery of different intracellular products. RESULTS: Here, we present a novel approach for extraction of integral membrane proteins from yeast based on cell envelope permeabilization through a combination of pulsed electric field and lytic enzyme pretreatment of the cells...
December 8, 2023: Microbial Cell Factories
https://read.qxmd.com/read/38055772/multiple-rounds-of-in-vivo-random-mutagenesis-and-selection-in-vibrio-natriegens-result-in-substantial-increases-in-ree-binding-capacity
#12
JOURNAL ARTICLE
Sean Medin, Anastacia Dressel, David A Specht, Timothy J Sheppard, Megan E Holycross, Matthew C Reid, Esteban Gazel, Mingming Wu, Buz Barstow
Rare earth elements (REE) are essential ingredients in many modern technologies, yet their purification remains either environmentally harmful or economically unviable. Adsorption, or biosorption, of REE onto bacterial cell membranes offers a sustainable alternative to traditional solvent extraction methods. But in order for biosorption-based REE purification to compete economically, the capacity and specificity of biosorption sites must be enhanced. Although there have been some recent advances in characterizing the genetics of REE-biosorption, the variety and complexity of bacterial membrane surface sites make targeted genetic engineering difficult...
December 6, 2023: ACS Synthetic Biology
https://read.qxmd.com/read/37930543/starting-with-an-integral-membrane-protein-project-for-structural-biology-production-purification-detergent-quantification-and-buffer-optimization-case-study-of-the-exporter-cnti-from-pseudomonas-aeruginosa
#13
JOURNAL ARTICLE
Maxime Mégret-Cavalier, Alexandre Pozza, Quentin Cece, Françoise Bonneté, Isabelle Broutin, Gilles Phan
Production, extraction, purification, and stabilization of integral membrane proteins are key steps for successful structural biology studies, in particular for X-ray crystallography or single particle microscopy. Here, we present the purification protocol of CntI from Pseudomonas aeruginosa, a new metallophore exporter of the Drug Metabolite Transporter (DMT) family involved in pseudopaline secretion. Subsequent to CntI purification, we optimized the buffer pH, salts, and additives by differential scanning fluorimetry (DSF), also known as Thermofluor Assay (TFA) or fluorescent thermal stability assay (FTSA), with the use of dye 1-AnilinoNaphthalene-8-Sulfonic acid (ANS), a fluorescent molecule compatible with detergents...
2024: Methods in Molecular Biology
https://read.qxmd.com/read/37877403/-production-and-antigenicity-analysis-of-a-recombinant-insulinoma-associated-protein-2-in-hek293-cells
#14
JOURNAL ARTICLE
Jingwen Qian, Pengbo Wang, Yuanxing Zhang, Qin Liu
Insulinoma-associated protein-2 (IA-2) is a transmembrane glycoprotein belonging to the tyrosine phosphatase-like protein family as well as an important autoantigen in the diagnosis of type 1 diabetes. IA-2 products have been marketed in Europe and the United States. At present, commercially available IA-2 antigens are either the recombinant IA-2ic domain or the IA-2 naturally extracted from bovine islets. However, the recombinant IA-2 antigen displays weak positive in clinic practice, which often results in occasional detection failures, thus cannot completely replace the naturally extracted IA-2 antigen...
October 25, 2023: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://read.qxmd.com/read/37875853/serodiagnosis-of-human-brucellosis-by-an-indirect-elisa-test-using-recombinant-outer-membrane-protein-19%C3%A2-kda-romp19-as-an-antigen
#15
JOURNAL ARTICLE
M Golchin, S Mollayi, E Mohammadi, N Eskandarzade
BACKGROUND: Brucellosis remains one of the global health concerns that reemerges in recent years. Delayed or inaccurate diagnosis end to a long treatment duration and financial burden; therefore, finding a good antigen for detection of specific anti-Brucella antibodies is crucial. We intended to evaluate the serodiagnosis value of recombinant Brucella outer membrane protein 19 kDa (rOMP19) using indirect ELISA system compared with Rose Bengal test. RESULTS: The OMP19 sequence was successfully cloned into pET-28a and produced in E...
October 24, 2023: BMC Biotechnology
https://read.qxmd.com/read/37851665/the-investigation-of-antibacterial-properties-of-peptides-and-protein-hydrolysates-derived-from-serum-of-asian-water-monitor-varanus-salvator
#16
JOURNAL ARTICLE
Jitkamol Thanasak, Sittiruk Roytrakul, Waraphan Toniti, Janthima Jaresitthikunchai, Narumon Phaonakrop, Siriwan Thaisakun, Sawanya Charoenlappanit, Rudee Surarit, Wanna Sirimanapong
It is well known that the Asian water monitors or Varanus salvator are both scavengers and predators. They can live and survive in the place that exposed to harmful microorganisms. Most people believe that they have some protected mechanisms to confront those infections. The aim of this study is to determine the antibacterial activities of crude peptides and protein hydrolysates extracted from serum of the Varanus salvator. Ten types of bacteria were cultured with crude peptides and protein hydrolysates which were isolated from 21 Varanus salvator's serum...
2023: PloS One
https://read.qxmd.com/read/37802583/aromatase-and-steroid-sulfatase-from-human-placenta
#17
JOURNAL ARTICLE
Debashis Ghosh
Cytochrome P450 aromatase (AROM) and steroid (estrone (E1)/dehydroepiandrosterone (DHEA)) sulfatase (STS) are the two key enzymes responsible for the biosynthesis of estrogens in human, and maintenance of the critical balance between androgens and estrogens. Human AROM, an integral membrane protein of the endoplasmic reticulum, is a member of the Fe-heme containing cytochrome P450 superfamily having a cysteine thiolate as the fifth Fe-coordinating ligand. It is the only enzyme known to catalyze the conversion of androgens with non-aromatic A-rings to estrogens characterized by the aromatic A-ring...
2023: Methods in Enzymology
https://read.qxmd.com/read/37719074/fast-and-sustainable-thermo-osmotic-dna-extraction-protocol-for-trans-spectrum-contingency-and-field-use
#18
JOURNAL ARTICLE
Stavroula Goudoudaki, Manousos E Kambouris, Marianna Manoussopoulou, George P Patrinos, Aristea Velegraki, Yiannis Manoussopoulos
In the field of molecular genetics, DNA extraction protocols and kits are sample-specific and proprietary, preventing lateral distribution among similar facilities from different sectors to alleviate supply shortages during a crisis. Expanding upon previous fast extraction protocols such as alkaline- and detergent-based ones, the use of boiling-hot water to rupture cells, virions, and nuclei, as proposed during the COVID-19 pandemic, might alleviate shortages and costs. Different soft, relatively abundant (highly enriched), and uncomplicated (genomically homogenous and with few inhibitors) biosamples are collected in 1...
September 5, 2023: Bio-protocol
https://read.qxmd.com/read/37665452/cell-surface-biotinylation-using-furan-cross-linking-chemistry
#19
JOURNAL ARTICLE
Esperanza Fernández, Laia Miret-Casals, Annemieke Madder, Kris Gevaert
A detailed study of the cellular surfaceome poses major challenges for mass spectrometry analysis. Surface proteins are low abundant compared to intracellular proteins, and their inefficient extraction in aqueous medium leads to their aggregation and precipitation. To tackle such problems, surface biotinylation is frequently used to tag surface proteins with biotin, allowing for their enrichment, leading to a more sensitive mapping of surface proteomes. We here detail a new surface biotinylation protocol based on furan-biotin affinity purification to enrich plasma membrane proteins for proteomics...
2023: Methods in Molecular Biology
https://read.qxmd.com/read/37647009/strategies-for-the-purification-of-membrane-proteins
#20
JOURNAL ARTICLE
Thomas J Butler, Sinéad Marian Smith
Membrane proteins account for approximately 30% of the coding regions of all sequenced genomes, and they play crucial roles in many fundamental cell processes. However, there are relatively few membrane proteins with known three-dimensional structures. This is likely due to technical challenges associated with membrane protein extraction, solubilization, and purification. Membrane proteins are classified based on the level of interaction with membrane lipid bilayers, with peripheral membrane proteins associating non-covalently with the membrane, and integral membrane proteins associating more strongly by means of hydrophobic interactions...
2023: Methods in Molecular Biology
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