488KB - The Doudna Lab - University of California, Berkeley
... Fitting the data to equation (2) yields a Km value of 2.8 mM for both products and a theoretical maximal velocity of 24 s–1 and 22 s–1 for the 11- or 12-nt and the 25- or 25-nt products, respectively. The maximum velocity (Vmax) of G. intestinalis Dicer is about ten-fold slower than that reported fo ...
... Fitting the data to equation (2) yields a Km value of 2.8 mM for both products and a theoretical maximal velocity of 24 s–1 and 22 s–1 for the 11- or 12-nt and the 25- or 25-nt products, respectively. The maximum velocity (Vmax) of G. intestinalis Dicer is about ten-fold slower than that reported fo ...
Nucleotide Metabolism
... Nucleotide Catabolism Begins with Nucleases to Release Nucleoside Monophosphates Nucleotidases Remove the Phosphate to Make Nucleosides Phosphorylates or Nucleosidases Release Bases and Sugars Adenine-Containing and Guanine Nucleotide Breakdown Processes are Similar, but Deaminases Co Inosine is Con ...
... Nucleotide Catabolism Begins with Nucleases to Release Nucleoside Monophosphates Nucleotidases Remove the Phosphate to Make Nucleosides Phosphorylates or Nucleosidases Release Bases and Sugars Adenine-Containing and Guanine Nucleotide Breakdown Processes are Similar, but Deaminases Co Inosine is Con ...
mineralization of chlorobenzene. of the meta
... been described. These organisms can be differentiated on the basis of the catabolic pathways dealing with the substituents. Chlorosubstituents can be removed by initial oxygenolytic, reductive, or hydrolytic reactions. Further mineralization can then occur via classical pathways such as the 3-oxoadi ...
... been described. These organisms can be differentiated on the basis of the catabolic pathways dealing with the substituents. Chlorosubstituents can be removed by initial oxygenolytic, reductive, or hydrolytic reactions. Further mineralization can then occur via classical pathways such as the 3-oxoadi ...
Microbial degradation of chloroaromatics Mars, Astrid E.
... Pseudomonas putida GJ31 is able to simultaneously grow on toluene and chlorobenzene. When cultures of this strain were inhibited with 3-fluorocatechol while growing on toluene or chlorobenzene, 3-methylcatechol or 3-chlorocatechol, respectively, accumulated in the medium. To establish the catabolic ...
... Pseudomonas putida GJ31 is able to simultaneously grow on toluene and chlorobenzene. When cultures of this strain were inhibited with 3-fluorocatechol while growing on toluene or chlorobenzene, 3-methylcatechol or 3-chlorocatechol, respectively, accumulated in the medium. To establish the catabolic ...
Biosynthesis of Glucosyl Glycerol, a Compatible Solute, Using
... that the suh gene from Xanthomonas axonopodis pv. glycines, a causative agent of bacterial pustule disease in soybeans, showed a very strong deduced amino acid homology with various ASases [17]. However, it only displayed sucrose hydrolysis activity without any glucosyltransferase or isomerization a ...
... that the suh gene from Xanthomonas axonopodis pv. glycines, a causative agent of bacterial pustule disease in soybeans, showed a very strong deduced amino acid homology with various ASases [17]. However, it only displayed sucrose hydrolysis activity without any glucosyltransferase or isomerization a ...
The semi-phosphorylative Entner–Doudoroff pathway in
... and subjected to a heat precipitation for 30 min at different temperatures. Extracts containing recombinant T. tenax and S. solfataricus protein were incubated at the following temperatures: T. tenax and S. solfataricus KD(P)G aldolase and T. tenax GAD at 85 ◦C (30 min), T. tenax GDH and KDG kinase ...
... and subjected to a heat precipitation for 30 min at different temperatures. Extracts containing recombinant T. tenax and S. solfataricus protein were incubated at the following temperatures: T. tenax and S. solfataricus KD(P)G aldolase and T. tenax GAD at 85 ◦C (30 min), T. tenax GDH and KDG kinase ...
Unusual ADP-forming acetyl-coenzyme A synthetases from the
... characterization of two ACDs with unique molecular and catalytic features, from the mesophilic euryarchaeon Haloarcula marismortui and from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum. ACD from H. marismortui was purified and characterized as a saltdependent, mesophilic ACD of homodimer ...
... characterization of two ACDs with unique molecular and catalytic features, from the mesophilic euryarchaeon Haloarcula marismortui and from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum. ACD from H. marismortui was purified and characterized as a saltdependent, mesophilic ACD of homodimer ...
GUIDE (download pdf)
... medium in which the target microorganisms grow and are detected by optically sensitive reagents. The reagents change their spectral patterns as the metabolic process takes place. These changes are detected photometrically by an optical instrument and monitored at predetermined time intervals. The ke ...
... medium in which the target microorganisms grow and are detected by optically sensitive reagents. The reagents change their spectral patterns as the metabolic process takes place. These changes are detected photometrically by an optical instrument and monitored at predetermined time intervals. The ke ...
Module 1. General principles of metabolism. Metabolism of
... C. * The pathway's end product represses the activity of the first enzyme in the pathway D. All of the above apply to feedback inhibition. E. More than one of the above, but not all, apply to feedback inhibition. 84. During electrophoretic research of LDG in the blood serum found out predominance of ...
... C. * The pathway's end product represses the activity of the first enzyme in the pathway D. All of the above apply to feedback inhibition. E. More than one of the above, but not all, apply to feedback inhibition. 84. During electrophoretic research of LDG in the blood serum found out predominance of ...
Unconventional serine proteases: Variations on the catalytic Ser/His
... Chymotrypsin- and trypsin-like proteases Two of the best-known serine proteases that utilize the Ser/His/Asp triad are chymotrypsin and trypsin. They share the same protein fold and have their catalytic residues in the order of His/Asp/Ser from the N to C terminus. Their active site regions are comp ...
... Chymotrypsin- and trypsin-like proteases Two of the best-known serine proteases that utilize the Ser/His/Asp triad are chymotrypsin and trypsin. They share the same protein fold and have their catalytic residues in the order of His/Asp/Ser from the N to C terminus. Their active site regions are comp ...
STRUCTURE-FUNCTION STUDIES OF THE CARNITINE/CHOLINE
... acid metabolism by facilitating the transfer of activated fatty acids across intracellular membranes. To understand the mechanism of these enzymes, the structure of human peroxisomal L-carnitine acetyltransferase (CAT) in complex with its substrate, L-carnitine, was determined. The structure reveale ...
... acid metabolism by facilitating the transfer of activated fatty acids across intracellular membranes. To understand the mechanism of these enzymes, the structure of human peroxisomal L-carnitine acetyltransferase (CAT) in complex with its substrate, L-carnitine, was determined. The structure reveale ...
Analysis of structural robustness of metabolic
... meaningful. Interestingly, if they are excluded, the robustness measure can never be exactly equal to one, since knockout of an enzyme then always deletes at least one elementary mode. In (1), no difference is made between different products of the metabolic network. For example, the contribution to ...
... meaningful. Interestingly, if they are excluded, the robustness measure can never be exactly equal to one, since knockout of an enzyme then always deletes at least one elementary mode. In (1), no difference is made between different products of the metabolic network. For example, the contribution to ...
The Biosynthesis of N-Phosphorylcreatine: an Investigation of the
... with the exception that ammediol buffer was used. Subsequently the following procedure was adopted. All operations were carried out at 2° and centrifuging was done 15 min. after the additions of (NH4)JSO. The muscle was treated for 2 min. in a Waring Blendor with 3 vol. of 0-01 Mammediol buffer (pH ...
... with the exception that ammediol buffer was used. Subsequently the following procedure was adopted. All operations were carried out at 2° and centrifuging was done 15 min. after the additions of (NH4)JSO. The muscle was treated for 2 min. in a Waring Blendor with 3 vol. of 0-01 Mammediol buffer (pH ...
This paper is published in a part-themed issue of Photochemical
... oxidation with release of AMP, pyrophosphate (PPi), CO2 and oxyluciferin, the light emitter. In parallel, D-LH2 –AMP is oxidized in a different way, giving rise to dehydroluciferyl–adenylate (L–AMP), which binds to luciferase inhibiting the light reaction. CoA can react with L–AMP giving rise to L–C ...
... oxidation with release of AMP, pyrophosphate (PPi), CO2 and oxyluciferin, the light emitter. In parallel, D-LH2 –AMP is oxidized in a different way, giving rise to dehydroluciferyl–adenylate (L–AMP), which binds to luciferase inhibiting the light reaction. CoA can react with L–AMP giving rise to L–C ...
Glycoside hydrolases: Catalytic base
... experiments are commonly used to identify catalytic residues. In a catalytic base mutant of an inverting GH, enzymatic activity is very low and it is not affected by substrates with different leaving groups. In contrast, removal of the acid residue inactivates hydrolysis of substrates with poor leav ...
... experiments are commonly used to identify catalytic residues. In a catalytic base mutant of an inverting GH, enzymatic activity is very low and it is not affected by substrates with different leaving groups. In contrast, removal of the acid residue inactivates hydrolysis of substrates with poor leav ...
Fulltext PDF - Indian Academy of Sciences
... were immunized using an enzyme in order to produce a monoclonal antibody (referred to as Ab1), the antigen-binding site of which is structurally complementary to the active site of the enzyme. Monoclonal antibodies (Ab2) specific for the antigenbinding site of the Ab1 are then subsequently generated ...
... were immunized using an enzyme in order to produce a monoclonal antibody (referred to as Ab1), the antigen-binding site of which is structurally complementary to the active site of the enzyme. Monoclonal antibodies (Ab2) specific for the antigenbinding site of the Ab1 are then subsequently generated ...
1. Sources of cholesterol • diet ~ 30 % • biosynthesis ~ 70% 2. Types
... Part A. Extraction of cholesterol from food. • Do all of Part A in the hood. • Split the combined supernatant into two centrifuge tubes. • #7 add 2.5 mL of 15% Na2SO4 into each of the centrifuge and spin. • You will need 4 centrifuge tubes total. • Water bath is setup in the hood. • You will need 9 ...
... Part A. Extraction of cholesterol from food. • Do all of Part A in the hood. • Split the combined supernatant into two centrifuge tubes. • #7 add 2.5 mL of 15% Na2SO4 into each of the centrifuge and spin. • You will need 4 centrifuge tubes total. • Water bath is setup in the hood. • You will need 9 ...
L02_2002
... • allosteric regulation of both GS & GP • substrate cycles • enzyme-catalyzed covalent modification of both GS &GP • covalent modifications are under hormonal control in the body, through their own enzymatic cascades In LIVER: Glycogen metabolism is ultimately controlled by GLUCAGON — a 29 amino aci ...
... • allosteric regulation of both GS & GP • substrate cycles • enzyme-catalyzed covalent modification of both GS &GP • covalent modifications are under hormonal control in the body, through their own enzymatic cascades In LIVER: Glycogen metabolism is ultimately controlled by GLUCAGON — a 29 amino aci ...
Lecture 2: Glycogen metabolism (Chapter 15)
... Why use glycogen to store energy rather than just using fat? (Since fat is more abundant than glycogen in the body and also stores energy) 1. Muscles "mobilize" (i.e., convert to energy) glycogen faster than fat. 2. Fatty acid residues cannot be metabolized anaerobically (that is, without oxygen). ...
... Why use glycogen to store energy rather than just using fat? (Since fat is more abundant than glycogen in the body and also stores energy) 1. Muscles "mobilize" (i.e., convert to energy) glycogen faster than fat. 2. Fatty acid residues cannot be metabolized anaerobically (that is, without oxygen). ...
Module 2 General principles of metabolism. Мetabolism of carbohy
... E. * All reactions produce some heat. 17. Active holoenzymes are formed from ____________ in the presence of _________. A. Cofactors; proteins B. Proteins; cofactors C. * Apoenzymes; cofactors D. Apoenzymes; proteins E. Apoenzymes; inactive holoenzymes 18. An allosteric inhibitor of an enzyme usual ...
... E. * All reactions produce some heat. 17. Active holoenzymes are formed from ____________ in the presence of _________. A. Cofactors; proteins B. Proteins; cofactors C. * Apoenzymes; cofactors D. Apoenzymes; proteins E. Apoenzymes; inactive holoenzymes 18. An allosteric inhibitor of an enzyme usual ...
148 - Blue Ridge Institute for Medical Research
... Protein kinases play a predominant regulatory role in nearly every aspect of cell biology and they can modify the function of a protein in almost every conceivable way. Protein phosphorylation can increase or decrease enzyme activity and it can alter other biological activities such as transcription ...
... Protein kinases play a predominant regulatory role in nearly every aspect of cell biology and they can modify the function of a protein in almost every conceivable way. Protein phosphorylation can increase or decrease enzyme activity and it can alter other biological activities such as transcription ...
Glycogen Metabolism
... The released Ca++ also activates Phosphorylase Kinase, which in muscle includes calmodulin as its d subunit. Phosphorylase Kinase is partly activated by binding of Ca++ to this subunit. ...
... The released Ca++ also activates Phosphorylase Kinase, which in muscle includes calmodulin as its d subunit. Phosphorylase Kinase is partly activated by binding of Ca++ to this subunit. ...
Structure of bacterial luciferase
... [3 subunit is folded alone, it forms a homodimer which is insensitive to proteases and does not unfold even after prolonged incubation in 5 M urea [39"'], conditions that would cause rapid unfolding of the native heterodimer [40-42]. These biochemical studies, both with the proteolyzed enzyine and w ...
... [3 subunit is folded alone, it forms a homodimer which is insensitive to proteases and does not unfold even after prolonged incubation in 5 M urea [39"'], conditions that would cause rapid unfolding of the native heterodimer [40-42]. These biochemical studies, both with the proteolyzed enzyine and w ...
Glycogen Metabolism
... The released Ca++ also activates Phosphorylase Kinase, which in muscle includes calmodulin as its d subunit. Phosphorylase Kinase is partly activated by binding of Ca++ to this subunit. ...
... The released Ca++ also activates Phosphorylase Kinase, which in muscle includes calmodulin as its d subunit. Phosphorylase Kinase is partly activated by binding of Ca++ to this subunit. ...
Kinetic studies of human tyrosyl
... and 0.125 lM enzyme with different concentrations of substrate ranging from 25 lM to 2000 lM. Increases in absorbance at 415 nm were monitored and the amount of released products was calculated as described above. The specific activity was determined as nmol of productÆmin)1Ælg)1 of enzyme. Km and kc ...
... and 0.125 lM enzyme with different concentrations of substrate ranging from 25 lM to 2000 lM. Increases in absorbance at 415 nm were monitored and the amount of released products was calculated as described above. The specific activity was determined as nmol of productÆmin)1Ælg)1 of enzyme. Km and kc ...
Enzyme inhibitor
An enzyme inhibitor is a molecule that binds to an enzyme and decreases its activity. Since blocking an enzyme's activity can kill a pathogen or correct a metabolic imbalance, many drugs are enzyme inhibitors. They are also used in pesticides. Not all molecules that bind to enzymes are inhibitors; enzyme activators bind to enzymes and increase their enzymatic activity, while enzyme substrates bind and are converted to products in the normal catalytic cycle of the enzyme.The binding of an inhibitor can stop a substrate from entering the enzyme's active site and/or hinder the enzyme from catalyzing its reaction. Inhibitor binding is either reversible or irreversible. Irreversible inhibitors usually react with the enzyme and change it chemically (e.g. via covalent bond formation). These inhibitors modify key amino acid residues needed for enzymatic activity. In contrast, reversible inhibitors bind non-covalently and different types of inhibition are produced depending on whether these inhibitors bind to the enzyme, the enzyme-substrate complex, or both.Many drug molecules are enzyme inhibitors, so their discovery and improvement is an active area of research in biochemistry and pharmacology. A medicinal enzyme inhibitor is often judged by its specificity (its lack of binding to other proteins) and its potency (its dissociation constant, which indicates the concentration needed to inhibit the enzyme). A high specificity and potency ensure that a drug will have few side effects and thus low toxicity.Enzyme inhibitors also occur naturally and are involved in the regulation of metabolism. For example, enzymes in a metabolic pathway can be inhibited by downstream products. This type of negative feedback slows the production line when products begin to build up and is an important way to maintain homeostasis in a cell. Other cellular enzyme inhibitors are proteins that specifically bind to and inhibit an enzyme target. This can help control enzymes that may be damaging to a cell, like proteases or nucleases. A well-characterised example of this is the ribonuclease inhibitor, which binds to ribonucleases in one of the tightest known protein–protein interactions. Natural enzyme inhibitors can also be poisons and are used as defences against predators or as ways of killing prey.