Cytoplasmic malate dehydrogenase from Phycomyces blakesleeanus: Kinetics and mechanism

1985 ◽  
Vol 63 (10) ◽  
pp. 1097-1105 ◽  
Author(s):  
Francisco Teixido ◽  
Dolores De Arriaga ◽  
Félix Busto ◽  
Joaquin Soler
Keyword(s):  
Malate Dehydrogenase ◽  
Ternary Complex ◽  
Initial Rate ◽  
Potassium Phosphate ◽  
Product Inhibition ◽  
Potassium Phosphate Buffer ◽  
Phycomyces Blakesleeanus ◽  
Coenzyme Binding ◽  
The Individual ◽  
Inhibition Studies

The kinetics and reaction mechanism of cytoplasmic malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37) from mycelium of Phycomyces blakesleeanus NRRL 1555 (−) in 0.1 M potassium phosphate buffer (pH 7.5) at 30 °C have been investigated. The initial rate and product inhibition studies were consistent with an ordered bi-bi mechanism that involved more than one kinetically significant ternary complex and also with the coenzyme binding first. The dissociation of the coenzyme from the enzyme–coenzyme complex appeared to be the slowest step in either direction of the reaction. The kinetic and rate constants for the individual steps of the reaction were determined.

scholarly journals Kinetic mechanism of Escherichia coli isocitrate dehydrogenase and its inhibition by glyoxylate and oxaloacetate

1986 ◽  
Vol 234 (2) ◽  
pp. 317-323 ◽  
Author(s):  
H G Nimmo
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Isocitrate Dehydrogenase ◽  
Initial Rate ◽  
Potent Inhibitor ◽  
Competitive Inhibition ◽  
Kinetic Mechanism ◽  
Product Inhibition ◽  
Coenzyme Binding ◽  
Inhibition Studies

The inhibition of Escherichia coli isocitrate dehydrogenase by glyoxylate and oxaloacetate was examined. The shapes of the progress curves in the presence of the inhibitors depended on the order of addition of the assay components. When isocitrate dehydrogenase or NADP+ was added last, the rate slowly decreased until a new, inhibited, steady state was obtained. When isocitrate was added last, the initial rate was almost zero, but the rate increased slowly until the same steady-state value was obtained. Glyoxylate and oxaloacetate gave competitive inhibition against isocitrate and uncompetitive inhibition against NADP+. Product-inhibition studies showed that isocitrate dehydrogenase obeys a compulsory-order mechanism, with coenzyme binding first. Glyoxylate and oxaloacetate bind to and dissociate from isocitrate dehydrogenase slowly. These observations can account for the shapes of the progress curves observed in the presence of the inhibitors. Condensation of glyoxylate and oxaloacetate produced an extremely potent inhibitor of isocitrate dehydrogenase. Analysis of the reaction by h.p.l.c. showed that this correlated with the formation of oxalomalate. This compound decomposed spontaneously in assay mixtures, giving 4-hydroxy-2-oxoglutarate, which was a much less potent inhibitor of the enzyme. Oxalomalate inhibited isocitrate dehydrogenase competitively with respect to isocitrate and was a very poor substrate for the enzyme. The data suggest that the inhibition of isocitrate dehydrogenase by glyoxylate and oxaloacetate is not physiologically significant.

scholarly journals Purification and kinetic properties of ox brain histamine N-methyltransferase

1986 ◽  
Vol 233 (3) ◽  
pp. 669-676 ◽  
Author(s):  
W L Gitomer ◽  
K F Tipton
Keyword(s):  
Acetic Acid ◽  
Steady State ◽  
Mouse Brain ◽  
Initial Rate ◽  
Substrate Inhibition ◽  
Gel Filtration ◽  
Product Inhibition ◽  
Kinetic Properties ◽  
Brain Histamine ◽  
Inhibition Studies

Histamine N-methyltransferase (EC 2.1.1.8) was purified 1100-fold from ox brain. The native enzyme has an Mr of 34800 +/- 2400 as measured by gel filtration on Sephadex G-100. The enzyme is highly specific for histamine. It does not methylate noradrenaline, adrenaline, DL-3,4-dihydroxymandelic acid, 3,4-dihydroxyphenylacetic acid, 3-hydroxytyramine or imidazole-4-acetic acid. Unlike the enzyme from rat and mouse brain, ox brain histamine N-methyltransferase did not exhibit substrate inhibition by histamine. Initial rate and product inhibition studies were consistent with an ordered steady-state mechanism with S-adenosylmethionine being the first substrate to bind to the enzyme and N-methylhistamine being the first product to dissociate.

scholarly journals Kinetic studies with the low-Km aldehyde reductase from ox brain

1985 ◽  
Vol 227 (2) ◽  
pp. 621-627 ◽  
Author(s):  
C M Ryle ◽  
K F Tipton
Keyword(s):  
Initial Velocity ◽  
Initial Rate ◽  
Kinetic Studies ◽  
Product Inhibition ◽  
Binary Complex ◽  
Aldehyde Reductase ◽  
Apparent Dissociation Constant ◽  
Displacement Mechanism ◽  
Sequential Mechanism ◽  
Inhibition Studies

Initial-rate studies of the low-Km aldehyde reductase-catalysed reduction of pyridine-3-aldehyde by NADPH gave families of parallel double-reciprocal plots, consistent with a double-displacement mechanism being obeyed. Studies on the variation of the initial velocity with the concentration of a mixture of the two substrates were also consistent with a double-displacement mechanism. In contrast, the initial-rate data indicated that a sequential mechanism was followed when NADH was used as the coenzyme. Product-inhibition studies, however, indicated that a compulsory-order mechanism was followed in which NADPH bound before pyridine-3-aldehyde with a ternary complex being formed and the release of pyrid-3-ylcarbinol before NADP+. The apparently parallel double-reciprocal plots obtained in the initial-rate studies with NADPH and pyridine-3-aldehyde were thus attributed to the apparent dissociation constant for the binary complex between the enzyme and coenzyme being finite but very low.

scholarly journals Kinetic studies on the reaction catalysed by phosphofructokinase from Trypanosoma brucei

1987 ◽  
Vol 245 (1) ◽  
pp. 13-18 ◽  
Author(s):  
C N Cronin ◽  
K F Tipton
Keyword(s):  
Trypanosoma Brucei ◽  
Initial Rate ◽  
Reaction Pathway ◽  
Potassium Phosphate ◽  
Kinetic Studies ◽  
Enzyme Concentration ◽  
Product Inhibition ◽  
Steady State Kinetics ◽  
Fructose 1,6 Bisphosphate ◽  
Kinetics Of

The steady-state kinetics of the reaction catalysed by the bloodstream form of Trypanosoma brucei were studied at pH 6.7. In the presence of 50 mM-potassium phosphate buffer, the apparent co-operativity with respect to fructose 6-phosphate and the non-linear relationship between initial velocity and enzyme concentration, which were found when the enzyme was assayed in 50 mM-imidazole buffer [Cronin & Tipton (1985) Biochem. J. 227, 113-124], are not evident. Studies on the variations of the initial rate with changing concentrations of MgATP and fructose 6-phosphate, the product inhibition by fructose 1,6-bisphosphate and the effects of the alternative substrate ITP were consistent with an ordered reaction pathway, in which MgATP binds to the enzyme before fructose 6-phosphate, and fructose 1,6-bisphosphate is the first product to dissociate from the ternary complex.

scholarly journals Drosophila melanogaster alcohol dehydrogenase: product-inhibition studies

1994 ◽  
Vol 301 (3) ◽  
pp. 901-909 ◽  
Author(s):  
J O Winberg ◽  
J S McKinley-McKee
Keyword(s):  
Acetic Acid ◽  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Ternary Complex ◽  
Product Inhibition ◽  
Ternary Complexes ◽  
Binary Complexes ◽  
Inhibition Studies ◽  
Inhibition Pattern ◽  
Drosophila Adh

The Drosophila melanogaster alleloenzymes AdhS and AdhF have been studied with respect to product inhibition by using the two substrate couples propan-2-ol/acetone and ethanol/acetaldehyde together with the coenzyme couple NAD+/NADH. With both substrate couples the reaction was consistent with an ordered Bi Bi mechanism. The substrates added to the enzyme in a compulsory order, with coenzyme as the leading substrate, to give two interconverting ternary complexes. The second ternary complex broke down with release of products in an obligatory order, with the aldehyde/ketone leaving first. Both the acetaldehyde and acetone products formed binary complexes with the enzyme that affected NAD+ binding. However, only an enzyme-acetone complex seemed to affect NADH binding and hence the reverse reaction. The inhibitory pattern with acetaldehyde as product was also affected by the formation of a ternary enzyme-NAD(+)-acetaldehyde complex, which broke down to acetic acid and NADH. The product-inhibition pattern shown in the present work is different from that published for Drosophila Adh previously and this discrepancy can not be explained by the use of different variants of Drosophila Adh.

Ultrastructure of Melanin Formation in Curvularia sp., Alternaria sp., and Drechslera Sorokiniana

1977 ◽  
Vol 35 ◽  
pp. 398-399
Author(s):  
M. H. Wheeler ◽  
W. J. Tolmsoff ◽  
A. A. Bell
Keyword(s):  
Potassium Phosphate ◽  
Thielaviopsis Basicola ◽  
Wild Type ◽  
Potassium Phosphate Buffer ◽  
Transmission Microscopy ◽  
Electron Transmission ◽  
Melanin Formation ◽  
Before And After ◽  
Alternaria Sp ◽  
Electron Transmission Microscopy

(+)-Scytalone [3,4-dihydro-3,6,8-trihydroxy-l-(2Hj-naphthalenone] and 1,8-di- hydroxynaphthalene (DHN) have been proposed as intermediates of melanin synthesis in the fungi Verticillium dahliae (1, 2, 3, 4) and Thielaviopsis basicola (4, 5). Scytalone is enzymatically dehydrated by V. dahliae to 1,3,8-trihydroxynaphthalene which is then reduced to (-)-vermelone [(-)-3,4- dihydro-3,8-dihydroxy-1(2H)-naphthalenone]. Vermelone is subsequently dehydrated to DHN which is enzymatically polymerized to melanin.Melanin formation in Curvularia sp., Alternaria sp., and Drechslera soro- kiniana was examined by light and electron-transmission microscopy. Wild-type isolates of each fungus were compared with albino mutants before and after treatment with 1 mM scytalone or 0.1 mM DHN in 50 mM potassium phosphate buffer, pH 7.0. Both chemicals were converted to dark pigments in the walls of hyphae and conidia of the albino mutants. The darkened cells were similar in appearance to corresponding cells of the wild types under the light microscope.

scholarly journals Product inhibition studies on bovine liver carbamoyl phosphate synthetase

1974 ◽  
Vol 141 (3) ◽  
pp. 817-824 ◽  
Author(s):  
Keith R. F. Elliott ◽  
Keith F. Tipton
Keyword(s):  
Inorganic Phosphate ◽  
Substrate Binding ◽  
Inorganic Ions ◽  
Bovine Liver ◽  
Product Inhibition ◽  
Carbamoyl Phosphate Synthetase ◽  
Carbamoyl Phosphate ◽  
Product Release ◽  
Proposed Model ◽  
Inhibition Studies

A study of the product-inhibition patterns of carbamoyl phosphate synthetase from bovine liver is reported. Inhibition by adenosine, AMP and inorganic ions is also reported. The results are in agreement with the previously proposed model in which the order of substrate binding is ATPMg, followed by HCO3−, ATPMg and NH4+. The order of product release on the basis of the reported results is carbamoyl phosphate, followed by ADPMg, ADPMg and inorganic phosphate.

scholarly journals Emergence of protocellular growth laws

2007 ◽  
Vol 362 (1486) ◽  
pp. 1841-1845 ◽  
Author(s):  
Tristan Rocheleau ◽  
Steen Rasmussen ◽  
Peter E Nielsen ◽  
Martin N Jacobi ◽  
Hans Ziock
Keyword(s):  
Kinetic Studies ◽  
Product Inhibition ◽  
Replication Process ◽  
Gene Replication ◽  
Math Biol ◽  
Curr Opin Cell Biol ◽  
Growth Laws ◽  
Internal Gene ◽  
The Many ◽  
The Individual

Template-directed replication is known to obey a parabolic growth law due to product inhibition (Sievers & Von Kiedrowski 1994 Nature 369 , 221; Lee et al . 1996 Nature 382 , 525; Varga & Szathmáry 1997 Bull. Math. Biol . 59 , 1145). We investigate a template-directed replication with a coupled template catalysed lipid aggregate production as a model of a minimal protocell and show analytically that the autocatalytic template–container feedback ensures balanced exponential replication kinetics; both the genes and the container grow exponentially with the same exponent. The parabolic gene replication does not limit the protocellular growth, and a detailed stoichiometric control of the individual protocell components is not necessary to ensure a balanced gene–container growth as conjectured by various authors (Gánti 2004 Chemoton theory ). Our analysis also suggests that the exponential growth of most modern biological systems emerges from the inherent spatial quality of the container replication process as we show analytically how the internal gene and metabolic kinetics determine the cell population's generation time and not the growth law (Burdett & Kirkwood 1983 J. Theor. Biol . 103 , 11–20; Novak et al . 1998 Biophys. Chem . 72 , 185–200; Tyson et al . 2003 Curr. Opin. Cell Biol . 15 , 221–231). Previous extensive replication reaction kinetic studies have mainly focused on template replication and have not included a coupling to metabolic container dynamics (Stadler et al . 2000 Bull. Math. Biol . 62 , 1061–1086; Stadler & Stadler 2003 Adv. Comp. Syst . 6 , 47). The reported results extend these investigations. Finally, the coordinated exponential gene–container growth law stemming from catalysis is an encouraging circumstance for the many experimental groups currently engaged in assembling self-replicating minimal artificial cells (Szostak 2001 et al . Nature 409 , 387–390; Pohorille & Deamer 2002 Trends Biotech . 20 123–128; Rasmussen et al . 2004 Science 303 , 963–965; Szathmáry 2005 Nature 433 , 469–470; Luisi et al . 2006 Naturwissenschaften 93 , 1–13). 1

scholarly journals Protecting ultra- and hyperhydrophilic implant surfaces in dry state from loss of wettability

2016 ◽  
Vol 2 (1) ◽  
pp. 557-560 ◽  
Author(s):  
Steffen Lüers ◽  
Markus Laub ◽  
Herbert P. Jennissen
Keyword(s):  
Dental Implants ◽  
Phosphate Buffer ◽  
Potassium Phosphate ◽  
Optimal System ◽  
Potassium Phosphate Buffer ◽  
Long Time ◽  
Optical Appearance

AbstractUltrahydrophilic titanium miniplates with sandblasted and acid etched (SLA) surfaces were protected from loss of hydrophilicity by an exsiccation layer of salt and stored in a dry state. Various salts in different concentrations were tested in respect to their conservation capacity and optical appearance. Potassium phosphate buffer in a specified composition appeared to be optimal. This optimal system was applied in a long time storage experiment showing no loss of hydrophilicity over years. It was also transferred with success to hyperhydrophilic dental implants.

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