Hemagglutininesterase is a protein of the envelope of some viruses. Its function is related with the pathogenicity of the virus and with its interaction with the host. It may help the virus bind and enter the mucus layer of the intestinal way.
UGT2B7 (UDP-Glucuronosyltransferase-2B7) is a phase II metabolism isoenzyme found to be active in the liver, kidneys, epithelial cells of the lower gastrointestinal tract and also has been reported in the brain.
This enzyme is located on the endoplasmic reticulum and nuclear membranes of cells. Its function is to catalyse the conjugation of a wide variety of lipophilic aglycon substrates with glucuronic acid, using uridine diphosphate glucuronic acid.
UGT2B7 is the major enzyme isoform for the metabolism of morphine to the main metabolites, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G).[1]
This gene encodes a UDP-glucuronosyltransferase, an enzyme of the glucuronidation pathway that transforms small lipophilic molecules, such as steroids, bilirubin, hormones, and drugs, into water-soluble, excretable metabolites. This gene is part of a complex locus that encodes several UDP-glucuronosyltransferases. The locus includes thirteen unique alternate first exons followed by four common exons. Four of the alternate first exons are considered pseudogenes. Each of the remaining nine 5′ exons may be spliced to the four common exons, resulting in nine proteins with different N-termini and identical C-termini. Each first exon encodes the substrate binding site, and is regulated by its own promoter. Mutations in this gene result in Crigler-Najjar syndromes types I and II and in Gilbert syndrome.[1]
Lack of expression of UGT1A1 in the neonatal liver is the major cause of jaundice in newborns. This jaundice is generally caused by the natural breakdown of fetal blood cells which produces bilirubin that cannot be cleared if UGT1A1 is expressed at low levels or is absent. This type of jaundice can remedied by UV light exposure.
Animals must metabolize proteins to amino acids, at the expense of muscle tissue, when blood sugar is low. The preference of liver transaminases for oxaloacetate or alpha-ketoglutarate plays a key role in funneling nitrogen from amino acid metabolism to Asp and Glu for conversion to urea for excretion of nitrogen. Similarly, in muscles the use of pyruvate for transamination gives Ala, which is carried by the bloodstream to the liver (the overall reaction being termed “glucose/alanine cycle”). Here other transaminases regenerate pyruvate, which provides a valuable precursor for gluconeogenesis. This alanine cycle is analogous to the Cori cycle which allows anaerobic metabolism by muscles.
In biochemistry, a transaminase or an aminotransferase is an enzyme that catalyzes a type of reaction between an amino acid and an ?-keto acid. Specifically, this reaction (transamination) involves removing the amino group from the amino acid, leaving behind an ?-keto acid, and transferring it to the reactant ?-keto acid and converting it into an amino acid. The enzymes are important in the production of various amino acids, and measuring the concentrations of various transaminases in the blood is important in the diagnosing and tracking many diseases. Transaminases require the coenzyme pyridoxal-phosphate, which is converted into pyridoxamine in the first phase of the reaction, when an amino acid is converted into a keto acid. Enzyme-bound pyridoxamine in turn reacts with pyruvate, oxaloacetate, or alpha-ketoglutarate, giving alanine, aspartic acid, or glutamic acid, respectively.
The presence of elevated transaminases can be an indicator of liver damage.
As mentioned above, sucrose phosphorylase is a very important enzyme in metabolism. The reaction catalyzed by sucrose phosphorylase produces the valuable byproducts alpha-D-glucose-1-phosphate and fructose. alpha-D-glucose-1-phosphate can be reversibly converted by phosphoglucomutase to glucose-6-phosphate (Tedokon et al. 1992), which is an important intermediate used in glycolysis. In addition, fructose can be reversibly converted into fructose-6-phosphate (Reid and Abratt 2005), also found in the glycolytic pathway. In fact, fructose-6-phosphate and glucose-6-phosphate can be interconverted in the glycolytic pathway by phosphohexose isomerase (Nelson and Cox 2005). The final product of glycolysis, pyruvate, has multiple implications in metabolism. During anaerobic conditions, pyruvate con be converted into either lactate or ethanol, depending on the organism, providing a quick source of energy. In aerobic conditions, pyruvate can be converted into Acetyl-CoA, which has many possible fates including catabolism in the Citric Acid Cycle for energy use and anabolism in the formation of fatty acids for energy storage. Through these reactions, sucrose phosphorylase becomes important in the regulation of metabolic functions.
The regulation of sucrose phosphorylase can also be used to explain its function in terms of energy consumption and preservation. The cAMP-CRP complex that enhances transcription of the sucrose phosphorylase gene (Reid and Abratt 2003) is only present when glucose levels are low. The purpose of sucrose phosphorylase, therefore, can be linked to the need for higher glucose levels, created through its reaction. The fact that glucose acts as a feedback inhibitor to prevent the formation of sucrose phosphorylase (Reid and Abratt 2005) further supports its catalytic role in the creation of glucose for energy use or storage.
The glucose-6-phosphate molecule created from the original alpha-D-glucose-1-phosphate product is also involved in the pentose phosphate pathway. Through a series of reactions, glucose-6-phosphate can be converted into ribose 5-phosphate, which is used for a variety of molecules such as nucleotides, coenzymes, DNA, and RNA (Nelson and Cox 2005). These connections reveal that sucrose phosphorylase is also important for the regulation of other cellular molecules.
Since the discovery and characterization of sucrose phosphorylase, few documented experiments discuss mechanisms of regulation for the enzyme. The known methods of regulation are transcriptional, affecting the amount of enzyme present at any given time.
Global regulation of DNA molecules containing the gene for sucrose phosphorylase is performed by [[repression|catabolite repression}. First discovered in Gram-negative bacteria, both Cyclic AMP (cAMP) and cAMP Receptor Protein (CRP) function in sucrose phosphorylase regulation (Reid and Abratt 2005). The cAMP-CRP complex formed when both molecules combine acts as a positive regulator for transcription of the sucrose phosphorylase gene. The complex binds to the promoter region to activate transcription, enhancing the creation of sucrose phosphorylase (Nelson and Cox 2005).
Genetic regulation of sucrose phosphorylase is also performed by metabolites. Through experimentation it is known that genes encoding for the sucrose phosphorylase enzyme can be induced by sucrose and raffinose (Trindade, Abratt, and Reid 2003). Glucose, on the other hand, represses the transcription of the sucrose phosphorylase gene (Trindade, Abratt, and Reid 2003). These metabolites undoubtedly function in this way because of their implications in cellular metabolism.
There has been little research on methods of the allosteric regulation of sucrose phosphorylase, so at this point the function of allosteric molecules can only be hypothesized. Due to the nature of its function in metabolic pathways, it is likely that sucrose phosphorylase is additionally regulated by other common metabolites. For example, the presence of ATP would probably inhibit sucrose phosphorylase since ATP is a product of the catabolic pathway. Conversely, ADP would likely stimulate sucrose phosphorylase to increase levels of ATP. Further research on the subject would be required to support or refute these ideas.
The structure of sucrose phosphorylase has been identified in numerous experiments. The enzyme consists of four major domains, namely A, B, B’, and C. Domains A, B’ and C exist as dimers around the active site (Sprogoe et al. 2004). The size of the enzyme, as determined by sedimentation centrifugation, was found to be 55 KDa, consisting of 488 amino acids (Koga et al. 1991). The active has been shown to contain two binding sites, one designated a water site where hydroxylic molecules such as 1,2-cyclohexanediol and ethylene glycol may bind, and another designated as the acceptor site where the sugar molecule binds. Though the function of the water site has not been completely elucidated, the enzyme’s stability in aqueous solutions indicates that the water site may be involved in hydrolysis of the glycosidic bond.
The acceptor site is surrounded by three active residues that have been found to be essential in enzymatic activity. Using specific mutagenic assays, Asp-192 was found to be the catalytic nucleophile of the enzyme, “attacking C-1 of the glucosyl moiety of sucrose” (Schwarz and Nidetzky 2006). In fact, in vitro manipulation has shown that D-xylose, L-sorbose, and L-arabinose can replace fructose as the glucosyl acceptor (Mieyal, Simon, and Abeles 1972). The only requirement of the acceptor molecule is that the hydroxyl group on the C-3 be cis-disposed to the oxygen atom of the glycosidic bond. Glu-232 acts as the Bronsted acid-base catalyst, donating a proton to the displaced hydroxyl group on C-1 of the glucoside (Schwarz, Brecker, and Nidetsky 2007).
The most significant residue in the enzymatic activity, however, is Asp-295 (Mueller and Nidetsky 2007). Upon cleavage of the fructofuranosyl moiety from sucrose, the resultant glucose forms a covalent intermediate with the enzyme. The carboxylate side chain of Asp-295 hydrogen bonds with the hydroxyl groups at C-2 and C-3 of the glucosyl residue (Mueller and Nidetsky 2007). This interaction is maximized during the transition state of this covalent complex, lending support to the ping-pong mechanism. Finally, phosphorylation of the glucosyl residue at C-1 forms a transient positive charge on the glucosyl carbon, promoting breakage of the ester bond between Asp-192 and the sugar residue (Schwarz and Nidetsky 2006). Cleavage yields the product, ?-D-glucose-1-phosphate.
The method by which sucrose phosphorylase converts sucrose to D-fructose and alpha-D-glucose-1-phosphate has been studied in great detail. In the reaction, sucrose is first phosphorylated, at which point fructose is released by the enzyme-substrate complex. A covalent glucose-enzyme complex results, with beta-linkage between an oxygen atom in the peptide bond and C-1 of glucose. The covalent complex was experimentally isolated by chemical modification of the protein using NaIO4 after addition of the substrate (Mirza et al. 2006), supporting the hypothesis that reaction catalyzed by sucrose phosphorylase proceeds through the ping-pong mechanism. In the final enzymatic step, the glycosidic bond is hydrolyzed through the promotion of a phosphate group, yielding alpha-D-glucose-1-phosphate.
In a separate reaction, alpha-D-glucose-1-phosphate is converted to glucose-6-phosphate by the action of phosphoglucomutase (Tedokon et al. 1992). Glucose-6-phosphate is an extremely important intermediate for several pathways in the human body, including glycolysis, gluconeogenesis, and the pentose phosphate pathway (Nelson and Cox 2005). The function of sucrose phosphorylase is especially significant due to the role alpha-D-glucose-1-phosphate in energy metabolism.
Sucrose phosphorylase (E.C. 2.4.1.7) is an important enzyme in the metabolism of sucrose and regulation of other metabolic intermediates. Sucrose phosphorylase is in the class of hexosyltransferases, a type of glycosyltransferase that catalyzes the transfer of a monosaccharide from a phosphorylated sugar molecule to an acceptor molecule. More specifically, sucrose phosphorylase catalyzes the conversion of sucrose to D-fructose and alpha-D-glucose-1-phosphate (Reid and Abratt 2005). It has been shown in multiple experiments that the enzyme catalyzes this conversion by a ping-pong mechanism.