• Genes
  • Commonly used reporter genes that induce visually identifiable characteristics usually involve fluorescent and luminescent proteins. (wikipedia.org)
  • Reporter genes can also be used to assay for the expression of the gene of interest, which may produce a protein that has little obvious or immediate effect on the cell culture or organism. (wikipedia.org)
  • A more complex use of reporter genes on a large scale is in two-hybrid screening, which aims to identify proteins that natively interact with one another in vivo. (wikipedia.org)
  • Proteins differ from one another primarily in their sequence of amino acids, which is dictated by the nucleotide sequence of their genes, and which usually results in protein folding into a specific three-dimensional structure that determines its activity. (wikipedia.org)
  • Residues
  • This has also explained the need for a thiol reagent like beta mercaptoethanol in the regeneration of the protein since such reagents weaken the sulfhydryl bonds between cysteine residues, expediting the regeneration of the aequorin. (wikipedia.org)
  • This reaction is important for chemical sequencing of proteins, as the Edman degradation process is unable to sequence more than 70 consecutive residues. (wikipedia.org)
  • Proteins (/ˈproʊˌtiːnz/ or /ˈproʊti.ɪnz/) are large biomolecules, or macromolecules, consisting of one or more long chains of amino acid residues. (wikipedia.org)
  • Short polypeptides, containing less than 20-30 residues, are rarely considered to be proteins and are commonly called peptides, or sometimes oligopeptides. (wikipedia.org)
  • The sequence of amino acid residues in a protein is defined by the sequence of a gene, which is encoded in the genetic code. (wikipedia.org)
  • Shortly after or even during synthesis, the residues in a protein are often chemically modified by post-translational modification, which alters the physical and chemical properties, folding, stability, activity, and ultimately, the function of the proteins. (wikipedia.org)
  • lipids
  • It is applicable to all kinds of kinase substrates regardless of their nature with no prior modification (peptides, protein, polymer, lipids, and sugars). (aacrjournals.org)
  • The assay is robust as indicated by the high Z' values (over 0.8), homogenous, can be completed in one step after completion of kinase reaction, does not require antibodies or custom synthesized substrates, and is ideal to search for optimal substrates in a collection of peptides, proteins, lipids in one assay format and to screen for ATP and Non-ATP competitive inhibitors. (aacrjournals.org)
  • Bacteria
  • A common reporter in bacteria is the E. coli lacZ gene, which encodes the protein beta-galactosidase. (wikipedia.org)
  • Similar pathways occur in other luminescent bacteria. (wikipedia.org)
  • His laboratory produced the first evidence for quorum sensing in bacteria, early evidence of the molecular mechanisms of circadian clock regulation in organisms (first using dinoflagellate luminescence and then expanded to other cellular proteins), and some of the initial studies of energy transfer in green fluorescent proteins (GFP) in cnidarian luminescence. (wikipedia.org)
  • Luminescent Bacteria: Hastings' investigations of luminous bacteria acted as a catalyst for the discoveries of the biochemical mechanisms involved in their light production, the discovery of a flavin to be a substrate in its luciferase reaction, the determination of gene regulation of the luciferases, and the first evidence for quorum sensing, a form of bacterial communication. (wikipedia.org)
  • peptides
  • The invention relates to the staining of poly(amino acids), including peptides, polypeptides and proteins in gels and on solid supports, using neutral or anionic complexes of transition metals. (google.com)
  • Humans
  • It is present in the milk of humans and other mammals, in the blood plasma and neutrophils and is one of the major proteins of virtually all exocrine secretions of mammals, such as saliva, bile, tears and pancreas. (wikipedia.org)
  • 1970s
  • Attention to lanthanide probes increased greatly in the mid-1970s when Finnish researchers proposed Eu(III), Sm(III), Tb(III), and Dy(III) polyaminocarboxylates as luminescent sensors in time-resolved luminescent (TRL) immunoassays. (wikipedia.org)
  • Molecule
  • Reporter gene assay have been increasingly used in high throughput screening (HTS) to identify small molecule inhibitors and activators of protein targets and pathways for drug discovery and chemical biology. (wikipedia.org)
  • They documented the molecular weight, isoelectric point, optical absorption spectra and presence of two iron atoms per protein molecule. (wikipedia.org)
  • Similarity in the size of exons and their distribution in the domains of the protein molecule indicates that the evolutionary development of lactoferrin gene occurred by duplication. (wikipedia.org)
  • aequorin
  • Chemical characterization of aequorin indicates the protein is somewhat resilient to harsh treatments. (wikipedia.org)
  • The early successful purification of aequorin led to the first experiments involving the injection of the protein into the tissues of living animals to visualize the physiological release of calcium in the muscle fibers of a barnacle. (wikipedia.org)
  • cells
  • The research, by scientists at McGill University in Montreal and the Radboud University Medical Center in the Netherlands, focuses on a complex of proteins, known as podosomes, found in the membrane of migratory cells and in certain invasive cancer cells. (mcgill.ca)
  • Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed monomeric fluorescent timer protein. (nih.gov)
  • Using a newly developed protein-based fluorescent timer, monomeric Kusabira Green Orange (mK-GO), which changes color with a predictable time course, here we show that small GTPase Rab27A effectors regulate age-dependent exocytosis of secretory vesicles in PC12 cells. (nih.gov)
  • HeLa cells were engineered to express Cx40-YFP or Q49X-YFP prior to immunolabeling for a resident protein of the Golgi apparatus (GM130) (A) or the ER (PDI) (B). Note that the mutant primarily colocalized with PDI. (nih.gov)
  • To determine the subcellular localizations of the mutant protein, Q49X-YFP-expressing HeLa cells were further immunolabeled with antibodies against either PDI, a resident protein of the ER, or GM130, a resident protein of the Golgi apparatus (Fig. 5). (nih.gov)
  • The benefit that GFP and other fluorescent proteins have over organic dyes or quantum dots is that they can be expressed exogenously in cells alone or as a fusion protein, a protein that is created by ligating the fluorescent gene (e.g. (wikipedia.org)
  • Lactoferrin is one of the transferrin proteins that transfer iron to the cells and control the level of free iron in the blood and external secretions. (wikipedia.org)
  • mEos fusion proteins allow for differential colour labelling in single cells, or groups of cells in developing organs. (wikipedia.org)
  • This mechanism allows for localized tagging of the protein and makes EosFP an appropriate tool for tracking protein movement within living cells. (wikipedia.org)
  • Like other fluorescent proteins, Eos can be used to report diverse signals in cells, tissues and organs without disturbing complex biological machinery. (wikipedia.org)
  • The novel property of photoinduced green-to-red conversion in Eos is useful because it allows for localized tracking of proteins in living cells. (wikipedia.org)
  • Chemical
  • it is the combined effect of all of the amino acid side chains in a protein that ultimately determines its three-dimensional structure and its chemical reactivity. (wikipedia.org)
  • Luciferase
  • Transcription of the LuxCDABEG operon results in luminescence due to the expression of LuxA and LuxB, which form a protein known as a luciferase and the expression of LuxC, D, E, and G, which are involved in the synthesis of the luciferase's substrate, tetradecanal. (wikipedia.org)
  • These structures correspond in size and location to the fluorescent bodies confirmed to be scintillons by their light emission, and they show colocalization of anti-luciferase and anti-LBP labeling meaning both bioluminescence proteins are found in the structures. (wikipedia.org)
  • Voltage gated proton channels were subsequently identified in a dinoflagellate confirming their predicted existence Scintillons have been extensively purified from L. polyedra by centrifugation, and these purified scintillon preparations contain luciferase and luciferin binding protein as the only detectable protein components. (wikipedia.org)
  • monomeric
  • Unlike the tetrameric fluorescent proteins derived from anthozoan coral, which can interfere with normal cellular function due to interactions between protein subunits, EosFP has been broken up into dimeric and monomeric variants through the introduction of single point mutations. (wikipedia.org)
  • luciferin
  • Unlike photoproteins which stably bind coelenterazine and emit light upon addition of calcium, coelenterazine is normally bound by RrLBP, the luciferin-binding protein. (wikipedia.org)
  • When synthesized, these two proteins aggregate together and migrate to the vacuole membrane where LBP binds luciferin and the scintillons acquires an ability to produce light upon stimulation. (wikipedia.org)
  • poorly
  • The first analyte is linked to a specific binding agent on a solid support such as a polymer and then another reaction couples the first poorly luminescent lanthanide complex with a new better one. (wikipedia.org)
  • fluorophores
  • Fluorophores can be attached to proteins via specific functional groups, such as amino groups (e.g. via succinimide, Isothiocyanate or hydrazine) carboxyl groups (e.g. via carbodiimide) thiol (e.g. via maleimide or acetyl bromide) azide (e.g. via click chemistry) or non-specificately (glutaraldehyde) or non-covalently (e.g. via hydrophobicity, etc. (wikipedia.org)
  • green
  • The blood of a cuttlefish is an unusual shade of green-blue, because it uses the copper-containing protein haemocyanin to carry oxygen instead of the red, iron-containing protein haemoglobin found in vertebrates' blood. (wikipedia.org)
  • While the main focus of his work was on the bioluminescence, Shimomura and two others, Martin Chalfie and Roger Tsien, were awarded the Nobel Prize in 2008 for their work on green fluorescent proteins. (wikipedia.org)
  • Some of this luminescent energy is transferred to the GFP, shifting the overall color towards green. (wikipedia.org)
  • All green-to-red photoinducible fluorescent proteins, including Eos, contain a chromophoric unit derived from the tripeptide his-tyr-gly. (wikipedia.org)
  • amino
  • In animals, proteins are needed in the diet to provide the essential amino acids that cannot be synthesized. (wikipedia.org)
  • Most proteins consist of linear polymers built from series of up to 20 different L-α-amino acids. (wikipedia.org)
  • Once linked in the protein chain, an individual amino acid is called a residue, and the linked series of carbon, nitrogen, and oxygen atoms are known as the main chain or protein backbone. (wikipedia.org)
  • The end with a free amino group is known as the N-terminus or amino terminus, whereas the end of the protein with a free carboxyl group is known as the C-terminus or carboxy terminus (the sequence of the protein is written from N-terminus to C-terminus, from left to right). (wikipedia.org)
  • reaction
  • Homoserine lactone is also a product of the proteolytic reaction of cyanogen bromide (CNBR) with a methionine residue in a protein. (wikipedia.org)
  • sensors
  • Combining luminescent atomically precise clusters with mesoflowers and nanofibres, he developed sensors at sub-zeptomole levels which are probably the limits of fast molecular detection. (wikipedia.org)
  • ions
  • Notably, the protein contains three EF hand motifs that function as binding sites for Ca2+ ions. (wikipedia.org)
  • Lanthanides can be used because their small size (ionic radius) gives them the ability to replace metal ions inside protein complex such as calcium or nickel. (wikipedia.org)
  • interact
  • In these cases it is important that both proteins be able to properly fold into their active conformations and interact with their substrates despite being fused. (wikipedia.org)
  • methods
  • Methods commonly used to study protein structure and function include immunohistochemistry, site-directed mutagenesis, X-ray crystallography, nuclear magnetic resonance and mass spectrometry. (wikipedia.org)
  • detection
  • Typically, expression levels of experimentally introduced proteins are not regulated, and high levels are often preferred to facilitate detection. (nasa.gov)
  • Subsequent time resolved detection of the metal-centered luminescent probe yields the desired signal. (wikipedia.org)
  • indicate
  • We also measure mRFP deactivation in solutions containing denaturants (guanidinium chloride or urea) or stabilizing co-solutes (glycerol or sucrose) frequently encountered in many protein formulations to test whether hydration of these co-solutes can be used to indicate their relative effects on protein kinetic stability. (biomedsearch.com)
  • Further colocalization experiments with the organelle residential proteins indicate that Q49X was retained in the endoplasmic reticulum. (nih.gov)
  • inhibitors
  • 0.8) , and rapid performance (10 minutes), and its ease of use, the luminescence based Kinase Glo TM, Kinase Glo Plus TM , and now Kinase Glo Max assay platform have gained wide acceptance in many drug screening programs for protein kinase inhibitors. (aacrjournals.org)
  • Since the linear range of ATP is extended to 500 µM, it is feasible to screen libraries for compounds that are not only competitive with ATP but also for those that are non competitive which broaden the selection of inhibitors of both serine/threonine protein kinases as well as tyrosine protein kinases. (aacrjournals.org)
  • backbone
  • Formation of the red chromophore involves cleaving the peptide backbone but includes almost no other changes in the protein structure. (wikipedia.org)
  • The other two dihedral angles in the peptide bond determine the local shape assumed by the protein backbone. (wikipedia.org)