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Enzyme Activity
Enzyme assays are laboratory methods for measuring enzyme, enzymatic activity. They are vital for the study of enzyme kinetics and enzyme inhibitor, enzyme inhibition. Enzyme units The quantity or concentration of an enzyme can be expressed in Mole (unit), molar amounts, as with any other chemical, or in terms of activity in enzyme units. Enzyme activity Enzyme activity is a measure of the quantity of active enzyme present and is thus dependent on various physical conditions, ''which should be specified''. It is calculated using the following formula: :\mathrm=\mathrm_\text=\mathrm\times\mathrm where :\mathrm = Enzyme activity :\mathrm_\text = Moles of substrate converted per unit time :\mathrm = Rate of the reaction :\mathrm = Reaction volume The SI unit is the katal, 1 katal = 1 mole (unit), mol s−1 (mole per second), but this is an excessively large unit. A more practical and commonly used value is enzyme unit (U) = 1 μmol min−1 (micromole per minute). 1 U correspon ...
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Stopped Flow
Stopped-flow is one of a number of methods of studying the kinetics of reactions in solution. It is ideal for studying chemical reactions with a typical dead time on the order of 1 millisecond. In the simplest form of the technique, the solutions of two reactants are rapidly mixed by being forced through a mixing chamber, on emerging from which the mixed fluid passes through an optical observation cell. At some point in time, the flow is suddenly stopped, and the reaction is monitored using a suitable spectroscopic probe, such as absorbance, Fluorescence spectroscopy, fluorescence or Fluorescence anisotropy, fluorescence polarization. The change in spectroscopic signal as a function of time is recorded, and the rate constants that define the reaction kinetics can then be obtained by fitting the data using a suitable model. Stopped-flow as an experimental technique was introduced by Britton Chance and extended by Quentin Gibson. Other techniques, such as the Temperature jump, temper ...
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Wavelength
In physics and mathematics, wavelength or spatial period of a wave or periodic function is the distance over which the wave's shape repeats. In other words, it is the distance between consecutive corresponding points of the same ''phase (waves), phase'' on the wave, such as two adjacent crests, troughs, or zero crossings. Wavelength is a characteristic of both traveling waves and standing waves, as well as other spatial wave patterns. The multiplicative inverse, inverse of the wavelength is called the ''spatial frequency''. Wavelength is commonly designated by the Greek letter lambda (''λ''). For a modulated wave, ''wavelength'' may refer to the carrier wavelength of the signal. The term ''wavelength'' may also apply to the repeating envelope (mathematics), envelope of modulated waves or waves formed by Interference (wave propagation), interference of several sinusoids. Assuming a sinusoidal wave moving at a fixed phase velocity, wave speed, wavelength is inversely proportion ...
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Fluorescence
Fluorescence is one of two kinds of photoluminescence, the emission of light by a substance that has absorbed light or other electromagnetic radiation. When exposed to ultraviolet radiation, many substances will glow (fluoresce) with colored visible light. The color of the light emitted depends on the chemical composition of the substance. Fluorescent materials generally cease to glow nearly immediately when the radiation source stops. This distinguishes them from the other type of light emission, phosphorescence. Phosphorescent materials continue to emit light for some time after the radiation stops. This difference in duration is a result of quantum spin effects. Fluorescence occurs when a photon from incoming radiation is absorbed by a molecule, exciting it to a higher energy level, followed by the emission of light as the molecule returns to a lower energy state. The emitted light may have a longer wavelength and, therefore, a lower photon energy than the absorbed radi ...
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Glucose-6-phosphate Dehydrogenase
Glucose-6-phosphate dehydrogenase (G6PD or G6PDH) () is a cytosolic enzyme that catalysis, catalyzes the chemical reaction : Glucose 6-phosphate, D-glucose 6-phosphate + NADP+ + 6-Phosphogluconolactone, 6-phospho-D-glucono-1,5-lactone + NADPH + H+ This enzyme participates in the pentose phosphate pathway (see image), a metabolic pathway that supplies reducing energy to cells (such as erythrocytes) by maintaining the level of the reduced form of the co-enzyme nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide phosphate (NADPH). The NADPH in turn maintains the level of glutathione in these cells that helps protect the red blood cells against oxidation, oxidative damage from compounds like hydrogen peroxide. Of greater quantitative importance is the production of NADPH for tissues involved in biosynthesis of fatty acids or isoprenoids, such as the liver, mammary glands, adipose tissue, and the adrenal glands. G6PD reduces NADP+ to NADPH while oxidizing glucose-6- ...
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Hexokinase
A hexokinase is an enzyme that irreversibly phosphorylates hexoses (six-carbon sugars), forming hexose phosphate. In most organisms, glucose is the most important substrate for hexokinases, and glucose-6-phosphate is the most important product. Hexokinase possesses the ability to transfer an inorganic phosphate group from ATP to a substrate. Hexokinases should not be confused with glucokinase, which is a specific hexokinase found in the liver. All hexokinases are capable of phosphorylating several hexoses but hexokinase IV(D) is often misleadingly called glucokinase, though it is no more specific for glucose than the other mammalian isoenzymes. Variation Genes that encode hexokinase have been discovered in every domain of life, and exist among a variety of species that range from bacteria, yeast, and plants to humans and other vertebrates. The enzymes from yeast, plants and vertebrates all show clear sequence evidence of homology, but those of bacteria may not be relat ...
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Coupled Assay
''Coupled'' is an American dating game show that aired on Fox from May 17 to August 2, 2016. It was hosted by television personality, Terrence J and created by Mark Burnett, of ''Survivor'', '' The Apprentice'', ''Are You Smarter than a 5th Grader?'', ''Shark Tank'', and '' The Voice'', as well as Ben Newmark, Dan Newmark and Larry Barron. Filming took place in Anguilla. The cast included Miss Arizona USA 2009, Alicia-Monique Blanco; Miss Colorado USA 2015, Talyah Polee; host, Domonique Price, and American singer-songwriter, TV personality, and former collegiate athlete; Alex Lagemann. Tyler Gattuso was in the running to be a cast member on '' Big Brother 17'', but was ultimately not chosen due to the news being leaked by online media and his Instagram hinting at the news suggesting he wasn't going to be active for quite a while. A few days later, after the cast for Big Brother was announced, he was ranting on Twitter, shortly deleting them and deactivating his account. On A ...
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Oxidoreductase
In biochemistry, an oxidoreductase is an enzyme that catalyzes the transfer of electrons from one molecule, the reductant, also called the electron donor, to another, the oxidant, also called the electron acceptor. This group of enzymes usually utilizes NADP+ or NAD+ as cofactors. Transmembrane oxidoreductases create electron transport chains in bacteria, chloroplasts and mitochondria, including respiratory complexes I, II and III. Some others can associate with biological membranes as peripheral membrane proteins or be anchored to the membranes through a single transmembrane helix.Superfamilies of single-pass transmembrane oxidoreductases
in Membranome database


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Redox
Redox ( , , reduction–oxidation or oxidation–reduction) is a type of chemical reaction in which the oxidation states of the reactants change. Oxidation is the loss of electrons or an increase in the oxidation state, while reduction is the gain of electrons or a decrease in the oxidation state. The oxidation and reduction processes occur simultaneously in the chemical reaction. There are two classes of redox reactions: * Electron transfer, Electron-transfer – Only one (usually) electron flows from the atom, ion, or molecule being oxidized to the atom, ion, or molecule that is reduced. This type of redox reaction is often discussed in terms of redox couples and electrode potentials. * Atom transfer – An atom transfers from one Substrate (chemistry), substrate to another. For example, in the rusting of iron, the oxidation state of iron atoms increases as the iron converts to an oxide, and simultaneously, the oxidation state of oxygen decreases as it accepts electrons r ...
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Nicotinamide Adenine Dinucleotide
Nicotinamide adenine dinucleotide (NAD) is a Cofactor (biochemistry), coenzyme central to metabolism. Found in all living cell (biology), cells, NAD is called a dinucleotide because it consists of two nucleotides joined through their phosphate groups. One nucleotide contains an adenine nucleobase and the other, nicotinamide. NAD exists in two forms: an Redox, oxidized and reduced form, abbreviated as NAD and NADH (H for hydrogen), respectively. In cellular metabolism, NAD is involved in redox reactions, carrying electrons from one reaction to another, so it is found in two forms: NAD is an oxidizing agent, accepting electrons from other molecules and becoming reduced; with H+, this reaction forms NADH, which can be used as a reducing agent to donate electrons. These electron transfer reactions are the main function of NAD. It is also used in other cellular processes, most notably as a substrate (biochemistry), substrate of enzymes in adding or removing chemical groups to or fr ...
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MTT Assay
The MTT assay is a colorimetric assay for assessing cell metabolic activity. NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present. These enzymes are capable of reducing the tetrazolium dye MTT, which is chemically 3-(4,5- di methyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its insoluble formazan, which has a purple color. Other closely related tetrazolium dyes including XTT, MTS and the WSTs, are used in conjunction with the intermediate electron acceptor, 1-methoxy phenazine methosulfate (PMS). With WST-1, which is cell-impermeable, reduction occurs outside the cell via plasma membrane electron transport. However, this traditionally assumed explanation is currently contended as proof has also been found of MTT reduction to formazan in lipidic cellular structures without apparent involvement of oxidoreductases. Tetrazolium dye assays can also be used to measure cytotoxicity (loss of viable cells) or ...
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