Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human

The development of fast and reliable methods for the identification of new bioactive compounds is of utmost importance to boost the process of drug discovery and development. Immobilized enzyme reactors (IMERs), integrated with high performance liquid chromatography (HPLC), are attractive and versatile tools for screening collections consisting of natural products and synthetic small molecules. Standard kinetic parameters of the immobilized enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from both Trypanosoma cruzi de and human have been determined (T. cruzi: K M G3P = 0.50 mmol L-1; K M NAD+ = 0.67 mmol L-1; humana: K M G3P = 3.7 mmol L-1; K M NAD+ = 0.75 mmol L-1), and comparisons of these values with those of the parasite and human free enzymes indicate a decrease in the affinity for the immobilized system (T. cruzi: K M G3P = 0.42 mmol L-1; K M NAD+ = 0.26 mmol L-1; humana: K M G3P = 0.16 mmol L-1; K M NAD+ = 0.18 mmol L-1). Interestingly, despite the kinetic differences between the two systems, the immobilized GAPDHs retained the required structural requirements for molecular recognition and biological activity, increasing the stability the enzyme. In the present work, we described an integrated structural analysis which has provided important insights into the molecular basis underlying the effects of immobilization on the ligand-receptor interactions and consequent enzymatic activity and kinetics parameters.

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Main Authors: Guido,Rafael V. C., Cardoso,Carmen L., Moraes,Marcela C. de, Andricopulo,Adriano D., Cass,Quezia B., Oliva,Glaucius
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Química 2010
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532010001000008
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spelling oai:scielo:S0103-505320100010000082010-12-01Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and humanGuido,Rafael V. C.Cardoso,Carmen L.Moraes,Marcela C. deAndricopulo,Adriano D.Cass,Quezia B.Oliva,Glaucius immobilized enzymes reactors enzymes kinetic parameters structural analysis drug design The development of fast and reliable methods for the identification of new bioactive compounds is of utmost importance to boost the process of drug discovery and development. Immobilized enzyme reactors (IMERs), integrated with high performance liquid chromatography (HPLC), are attractive and versatile tools for screening collections consisting of natural products and synthetic small molecules. Standard kinetic parameters of the immobilized enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from both Trypanosoma cruzi de and human have been determined (T. cruzi: K M G3P = 0.50 mmol L-1; K M NAD+ = 0.67 mmol L-1; humana: K M G3P = 3.7 mmol L-1; K M NAD+ = 0.75 mmol L-1), and comparisons of these values with those of the parasite and human free enzymes indicate a decrease in the affinity for the immobilized system (T. cruzi: K M G3P = 0.42 mmol L-1; K M NAD+ = 0.26 mmol L-1; humana: K M G3P = 0.16 mmol L-1; K M NAD+ = 0.18 mmol L-1). Interestingly, despite the kinetic differences between the two systems, the immobilized GAPDHs retained the required structural requirements for molecular recognition and biological activity, increasing the stability the enzyme. In the present work, we described an integrated structural analysis which has provided important insights into the molecular basis underlying the effects of immobilization on the ligand-receptor interactions and consequent enzymatic activity and kinetics parameters.info:eu-repo/semantics/openAccessSociedade Brasileira de QuímicaJournal of the Brazilian Chemical Society v.21 n.10 20102010-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532010001000008en10.1590/S0103-50532010001000008
institution SCIELO
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country Brasil
countrycode BR
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access En linea
databasecode rev-scielo-br
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region America del Sur
libraryname SciELO
language English
format Digital
author Guido,Rafael V. C.
Cardoso,Carmen L.
Moraes,Marcela C. de
Andricopulo,Adriano D.
Cass,Quezia B.
Oliva,Glaucius
spellingShingle Guido,Rafael V. C.
Cardoso,Carmen L.
Moraes,Marcela C. de
Andricopulo,Adriano D.
Cass,Quezia B.
Oliva,Glaucius
Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
author_facet Guido,Rafael V. C.
Cardoso,Carmen L.
Moraes,Marcela C. de
Andricopulo,Adriano D.
Cass,Quezia B.
Oliva,Glaucius
author_sort Guido,Rafael V. C.
title Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
title_short Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
title_full Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
title_fullStr Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
title_full_unstemmed Structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi and human
title_sort structural insights into the molecular basis responsible for the effects of immobilization on the kinetic parameters of glyceraldehyde-3-phosphate dehydrogenase from trypanosoma cruzi and human
description The development of fast and reliable methods for the identification of new bioactive compounds is of utmost importance to boost the process of drug discovery and development. Immobilized enzyme reactors (IMERs), integrated with high performance liquid chromatography (HPLC), are attractive and versatile tools for screening collections consisting of natural products and synthetic small molecules. Standard kinetic parameters of the immobilized enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from both Trypanosoma cruzi de and human have been determined (T. cruzi: K M G3P = 0.50 mmol L-1; K M NAD+ = 0.67 mmol L-1; humana: K M G3P = 3.7 mmol L-1; K M NAD+ = 0.75 mmol L-1), and comparisons of these values with those of the parasite and human free enzymes indicate a decrease in the affinity for the immobilized system (T. cruzi: K M G3P = 0.42 mmol L-1; K M NAD+ = 0.26 mmol L-1; humana: K M G3P = 0.16 mmol L-1; K M NAD+ = 0.18 mmol L-1). Interestingly, despite the kinetic differences between the two systems, the immobilized GAPDHs retained the required structural requirements for molecular recognition and biological activity, increasing the stability the enzyme. In the present work, we described an integrated structural analysis which has provided important insights into the molecular basis underlying the effects of immobilization on the ligand-receptor interactions and consequent enzymatic activity and kinetics parameters.
publisher Sociedade Brasileira de Química
publishDate 2010
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532010001000008
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