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  • eukaryotic
  • Methods for recombinant production of eukaryotic membrane proteins, yielding sufficient quantity and quality of protein for structural biology, remain a challenge. (jove.com)
  • This work provides support for the use of Saccharomyces cerevisiae as a suitable expression system for recombinant production of multi-domain eukaryotic membrane proteins. (jove.com)
  • membrane protein complexes: photosystem I, ATP synthase large DNA and protein complexes: nucleosome centriole and microtubule-organizing center (MTOC) cytoskeleton flagellum nucleolus stress granule germ cell granule neuronal transport granule Eukaryotic cells are structurally complex, and by definition are organized, in part, by interior compartments that are themselves enclosed by lipid membranes that resemble the outermost cell membrane. (wikipedia.org)
  • Heme-containing transmembrane ferric reductase domains (FRD) are found in both bacterial and eukaryotic proteins including ferric reductases (FRE), and NADPH oxidases (NOX). (wikipedia.org)
  • purification
  • Therefore, only a small proportion of MTBs can be axenically isolated and cultured, and the obtainable cell densities do not allow the purification of sufficient numbers of magnetosomes for industrial applications ( Jogler and Schüler, 2009 ). (frontiersin.org)
  • Biotinylated protein was detected using specific antibody directed against SERCA2 and, advantageously, GFP-His8 fusion protein was easily traced during the purification steps using in-gel fluorescence. (jove.com)
  • Importantly, talon resin affinity purification proved more specific than Ni-NTA resin for the GFP-His8 tagged protein, providing better separation of oligomers present, during size exclusion chromatography. (jove.com)
  • Protein purification protocols should combine efficiency, simplicity and cost effectiveness 1 . (jove.com)
  • Here, we describe the GST-His method as a new small-scale affinity purification system for recombinant proteins, based on a N-terminal Glutathione Sepharose Tag (GST) 2,3 and a C-terminal 10xHis tag 4 , which are both fused to the protein of interest. (jove.com)
  • Purification of magnetosomes is accomplished by use of a magnetic separation column after disruption of the cell membrane. (wikipedia.org)
  • fusion proteins
  • Two different expression vectors were utilised, allowing expression of C-terminal fusion proteins with a biotinylation domain or a GFP- His8 tag. (jove.com)
  • family proteins
  • The crystal structure of smu.1377c, a hypothetical protein from S. mutans , shows a similar fold to Sua5_YciO_YrdC-family proteins and indicates its functional role in tRNA modification. (iucr.org)
  • biological
  • Harnessing these proteins presents a biological (ambient condition) synthetic approach to producing tailored MNPs. (biochemsoctrans.org)
  • Crystal and solution structures of Rv1848 protein and their implications in the biological assembly of Mtb urease is presented. (iucr.org)
  • prokaryotic
  • We show that magnetite forms through phase transformation from a highly disordered phosphate-rich ferric hydroxide phase, consistent with prokaryotic ferritins, via transient nanometric ferric (oxyhydr)oxide intermediates within the magnetosome organelle. (pnas.org)
  • physiological
  • To increase cell density and magnetosome production, it is necessary to elucidate the physiological characteristics of a particular strain during cell growth and develop an appropriate artificial control strategy. (frontiersin.org)
  • However, it might influence physiological properties of the protein. (jove.com)
  • It has been proposed that under normal physiological conditions, these proteins may function by essentially the same mechanism. (wikipedia.org)
  • metabolism
  • Quantitative real-time reverse transcriptase PCR indicated that not only the expression of pdh operon but also genes related to magnetosomes biosynthesis and tricarboxylic acid cycle decreased dramatically, suggesting a link between carbon metabolism and magnetosome formation. (frontiersin.org)
  • These non-ideal arrangements may be pointing to additional, currently unknown functions of magnetosomes, possibly related to metabolism. (wikipedia.org)
  • biochemical
  • Key assays in enzymology for the biochemical characterization of proteins in vitro necessitate high concentrations of the purified protein of interest. (jove.com)
  • concentrations
  • MamC-GFP-tagged magnetosomes purified from cells displayed strong fluorescence, which was sensitive to detergents but stable under a wide range of temperature and salt concentrations. (environmental-expert.com)
  • In prokaryotes, the primary function of the cell wall is to protect the cell from internal turgor pressure caused by the much higher concentrations of proteins and other molecules inside the cell compared to its external environment. (wikipedia.org)
  • cell
  • The key time point for cell growth and magnetosome formation was found to be 18-20 h. (frontiersin.org)
  • At this point, cells entered the log phase of growth, the maximal values of C mag (1.78), iron content (0.47%), and magnetosome number (26 ± 3 per cell) were observed, superoxide dismutase (SOD) activity began to decrease more rapidly, ATP content dropped to an extremely low level (0.17 fmol), and reducing power (NADH/NAD + ratio) began to increase very rapidly. (frontiersin.org)
  • At the key time point (18-20 h), cell density is high and magnetosomes have matured. (frontiersin.org)
  • Culture conditions that effectively minimize dissolved oxygen accumulation, lactic acid content, and reducing power at this point will enhance magnetosome yield without obvious inhibition of cell growth. (frontiersin.org)
  • Improved methods are needed to increase MTB cell density and magnetosome yield at a reasonable cost. (frontiersin.org)
  • These proteins are important for cell survival during exposure to toxic metabolites, possibly because they can release K+, allowing H+ uptake. (wikipedia.org)
  • efflux
  • Among the functionally well-characterized members of the family are: KefB/KefC K+ efflux proteins of E. coli (i.e. (wikipedia.org)
  • species
  • These proteins, specific to MTB, are designated with the prefix Mam or Mms, although some are not found in every species of MTB. (frontiersin.org)
  • Cryptochrome, a flavoprotein found in the eyes of European robins and other animal species, is the only protein known to form photoinduced radical-pairs in animals. (wikipedia.org)
  • cells
  • The latter construct is used to generate baculoviruses, for infection of Sf9 infected cells for protein expression 5 . (jove.com)
  • One reported example includes large (up to 200 nm) magnetosomes found in coccoid cells in Brazil. (wikipedia.org)
  • minerals
  • Several other iron sulfide minerals have also been identified in iron sulfide magnetosomes-including mackinawite (tetragonal FeS) and a cubic FeS-which are thought to be precursors of Fe3S4. (wikipedia.org)
  • full length protein
  • The data were collected on the full length protein. (predictioncenter.org)
  • In summary, the combination of two different tags flanking the N- and the C-terminal and the capability to cleave off one of the tags, guaranties the recovery of a highly purified and full-length protein of interest. (jove.com)
  • iron
  • From the evidence, we propose a mechanism for its function as a specific magnetite nucleation protein and summaries the key features for this action: namely, self-assembly to display a charged surface for specific iron binding, with the curvature of the surfaces determining the particle size. (biochemsoctrans.org)
  • Synthetic Analogs of the Active Sites of Iron-Sulfur Proteins. (wikipedia.org)
  • purification
  • This step is crucial because it allows us to determine how our mutant compares with the wild-type on a quantitative level, as high activity without purification could simply be the result of high protein concentration. (igem.org)
  • Therefore, only a small proportion of MTBs can be axenically isolated and cultured, and the obtainable cell densities do not allow the purification of sufficient numbers of magnetosomes for industrial applications ( Jogler and Schüler, 2009 ). (frontiersin.org)
  • transmembrane
  • The proteins of the CPA2 family consist of between 333 and 900 amino acyl residues and exhibit 10-14 transmembrane α-helical spanners (TMSs). (wikipedia.org)
  • Each of these proteins consists of a transmembrane hydrophobic N-terminal domain, and a lesser conserved C-terminal hydrophilic domain. (wikipedia.org)
  • YedZ homologues can function as heme-binding proteins that facilitate or regulate oxidoreduction, transmembrane electron flow and transport. (wikipedia.org)
  • NOX enzymes are distinguished from FRE enzymes through a four amino acid motif spanning from transmembrane domain 3 (TM3) to TM4, and YedZ/STEAP proteins are identified by the replacement of the first canonical heme-spanning histidine by a highly conserved arginine. (wikipedia.org)
  • Steap family proteins are defined by a shared transmembrane domain that in Steap3 has been shown to function as a transmembrane electron shuttle, moving cytoplasmic electrons derived from NADPH across the lipid bilayer to the extracellular face where they are used to reduce Fe3+ to Fe2+ and potentially Cu2+ to Cu1+. (wikipedia.org)
  • scaffold
  • The variable loop of the peptide aptamer is generated through a random nucleotide library and held together by an optimized, enzymatic inactive version of human Thioredoxin (hTrx) used as a scaffold protein. (diyhpl.us)
  • coli
  • His work focused on the development for the first time of a method to directly quantify IPTG in E. coli cytoplasm and medium samples using LC-MS analysis and this methodology led to a better knowledge and understanding of inducible systems, gene regulation and optimization of recombinant protein production. (aston.ac.uk)
  • Among the functionally well-characterized members of the family are: KefB/KefC K+ efflux proteins of E. coli (i.e. (wikipedia.org)
  • cell
  • The key time point for cell growth and magnetosome formation was found to be 18-20 h. (frontiersin.org)
  • At this point, cells entered the log phase of growth, the maximal values of C mag (1.78), iron content (0.47%), and magnetosome number (26 ± 3 per cell) were observed, superoxide dismutase (SOD) activity began to decrease more rapidly, ATP content dropped to an extremely low level (0.17 fmol), and reducing power (NADH/NAD + ratio) began to increase very rapidly. (frontiersin.org)
  • At the key time point (18-20 h), cell density is high and magnetosomes have matured. (frontiersin.org)
  • Culture conditions that effectively minimize dissolved oxygen accumulation, lactic acid content, and reducing power at this point will enhance magnetosome yield without obvious inhibition of cell growth. (frontiersin.org)
  • Improved methods are needed to increase MTB cell density and magnetosome yield at a reasonable cost. (frontiersin.org)
  • cellular penetration capability protein using Clostridium difficile toxin B (tcdB), for the delivery of Cas9-gRNA into a cell for genome editing "C. diff toxin B (tcdB) is a marvelously evolved protein, dedicated to crossing cell membranes. (diyhpl.us)
  • These proteins are important for cell survival during exposure to toxic metabolites, possibly because they can release K+, allowing H+ uptake. (wikipedia.org)
  • complexes
  • At every point along the way, multifunctional proteins and [ribonucleoprotein] complexes facilitate communication between upstream and downstream steps, providing both feedforward and feedback information essential for proper coordination of what can only be described as an intricate and astonishing web of regulation. (icr.org)
  • species
  • Cryptochrome, a flavoprotein found in the eyes of European robins and other animal species, is the only protein known to form photoinduced radical-pairs in animals. (wikipedia.org)
  • interactions
  • Previous studies showed that DNA methylation helps cells with "gene regulation, DNA-protein interactions, cellular differentiation, suppression of transposable elements, embryogenesis, X-inactivation, genomic imprinting and tumorigenesis. (icr.org)
  • cells
  • The process of magnetosome synthesis involves a high consumption of ATP and reducing power, and the cells require replenishment of nutrients prior to the 18-20 h time point. (frontiersin.org)
  • One reported example includes large (up to 200 nm) magnetosomes found in coccoid cells in Brazil. (wikipedia.org)
  • iron
  • Synthetic Analogs of the Active Sites of Iron-Sulfur Proteins. (wikipedia.org)
  • Several other iron sulfide minerals have also been identified in iron sulfide magnetosomes-including mackinawite (tetragonal FeS) and a cubic FeS-which are thought to be precursors of Fe3S4. (wikipedia.org)
  • formation
  • The identification of these proteins as members of the CPA2 family reveals that monovalent cation transport is required for Bacillus spore formation and germination. (wikipedia.org)
  • toxin
  • A new study published in the journal Nature showed that the Photorhabdus bacteria's protein toxin, the 'ABC toxin,' is precisely engineered. (icr.org)
  • applications
  • Development of bioprocess for recombinant protein production in a wide range of hosts for biotech and healthcare applications. (aston.ac.uk)
  • point
  • In order to design mutations to wild-type Kumamolisin that would increase the enzyme's proteolytic activity on gluten, we used a computational enzyme editing program called Foldit, which allows the user to hypothetically modify the amino acid sequence of a protein by creating point mutations at any location within the protein's crystal structure. (igem.org)
  • conditions
  • It has been postulated that the evolutionary advantage of possessing a system of magnetosomes is linked to the ability of efficiently navigating within this zone of sharp chemical gradients by simplifying a potential three-dimensional search for more favourable conditions to a single dimension (see the "Magnetism" subsection below for a description of this mechanism). (wikipedia.org)
  • present
  • The polysaccharides, proteins and lipids present in the algal membranes act as capping agents and thus limit the use of non-biodegradable commercial surfactants, which are difficult to remove after the synthesis of MNPs. (scientiaricerca.com)
  • technology
  • The layout and construction of novel peptides and proteins occupy pivotal positions in technology and expertise and should proceed to take action within the twenty first century. (croustiglam.com)