Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana

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Main Author: Barbosa, Emmanuel Duarte
Other Authors: 91206359072
Language:Portuguese
Published: PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA 2017
Subjects:
DFT
Online Access:https://repositorio.ufrn.br/jspui/handle/123456789/21908
id ndltd-IBICT-oai-repositorio.ufrn.br-123456789-21908
record_format oai_dc
collection NDLTD
language Portuguese
sources NDLTD
topic CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
ALAD
Energia de liga??o
MFCC
DFT
spellingShingle CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
ALAD
Energia de liga??o
MFCC
DFT
Barbosa, Emmanuel Duarte
Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
description Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-02-02T13:30:50Z No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) === Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-02-08T19:26:36Z (GMT) No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) === Made available in DSpace on 2017-02-08T19:26:36Z (GMT). No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) Previous issue date: 2016-07-29 === A enzima Delta Aminolevul?nico Desidratase (ALAD) ? uma metaloprote?na citos?lica essencial em v?rios processos biol?gicos, uma vez que ? respons?vel pelo segundo passo da cat?lise enzim?tica na forma??o de porfobilinog?nio, um precursor dos tetrapirr?licos (heme, clorofila). Esta enzima ? bastante sens?vel a metais pesados e tem sido classicamente usada como um marcador na intoxica??o por chumbo. Sua inibi??o se d? pela substitui??o desses metais pesados no s?tio de liga??o a metais. Na ALAD humana, o Zinco (Zn2+) ocupa funcionalmente este s?tio sendo essencial para a coordena??o das cadeias de ?cido aminolevul?nico durante a cat?lise enzim?tica. Embora muitos ensaios in vitro, in vivo e in s?lico j? tenham demonstrado a import?ncia do Zn2+ nesse s?tio, n?o se tinha conhecimento de nenhum estudo baseado em abordagem qu?ntica com o intuito de elucidar esta intera??o de forma mais detalhada. Diante disso, o presente trabalho teve como objetivo analisar as muta??es missense que acometem o s?tio de liga??o ao zinco e descrever atrav?s de m?todos qu?nticos a energia de intera??o entre a enzima e o zinco com maior acur?cia utilizando o m?todo do Fracionamento Molecular com Capas Conjugadas (MFCC), quantificando energeticamente os res?duos de amino?cidos posicionados at? uma dist?ncia de 8,5 ? do centroide do ligante. Foi identificado as altera??es bioqu?micas na estrutura monom?rica dos mutantes, as quais resultam na diminui??o da atividade enzim?tica. Foram identificados um total de 30 res?duos com valores energ?ticos variados que interagem com o zinco no bols?o de liga??o. Aqueles que apresentaram valores significativos (de atra??o ou repuls?o) e est?o relacionados funcionalmente ? atividade enzim?tica foram: Lis199, Lis252, Arg 209, Arg 174, Cis122, Cis124 e Cis132; e aqueles que demonstraram relev?ncia para a perman?ncia do ?on no s?tio de liga??o foram: Asp169, Gli130, Gli133, Asp120 e Ser168. A partir disso, p?de-se concluir que al?m dos grupos nucle?filos (grupos tiolatos) dos res?duos Cis122, Cis124 e Cis132, os res?duos Asp169, Asp120 e Ser168 s?o fundamentais na composi??o do bols?o, uma vez que demonstraram grande quantidade de energia de intera??o atrativa com o ?on Zn2+. === The enzyme Delta Aminolevulinic Dehydratase (ALAD) is a cytosolic metalloproteinase essential in several biological processes since it participates in the second step in porphobilinogen formation pathway, a tetrapyrrolic precursor of heme and chlorophyll. This enzyme is very sensitive to heavy metals and has traditionally been used as a biomarker in lead poisoning. Its inhibition occurs when these heavy metals are replaced inside the metal binding site. In human ALAD, Zinc (Zn2+) functionally occupies this site and it is essential for coordination of two chains of aminolevulinic acid for the enzymatic catalysis. Although many in vitro, in vivo and in silico works have already demonstrated the importance of Zn2+ at that site, to the best of our knowledge, there isn?t any studies on literature based on quantum approach in order to elucidate this interactions in more details. Therefore, the aim of the present study was to analyse the missense mutations that affect the zinc binding site and describe through quantum methods the energy interaction between zinc and ALAD with greater accuracy using the method of Molecular fractionation with conjugated caps (MFCC) by quantifying amino acid residues? energy positioned at 8.5 ? of distance with the ligand centroid. It was identified biochemical changes in the monomeric structure of mutants, which result in decreased enzyme activity. It were identified a total of 30 residues with a wide range of energy values. The residues with significant (atractition or repulsion) values and functionally related to enzymatic activity were: Lys199, Lys252, Cys122, Cys124 and Cys132; and those that demonstrated relevance to the ion permanence inside the binding site were: Asp169, Gly130, Gly133, Asp120 and Ser168. Thus, it could be concluded that in addition to the nucleophilic groups (thiolates groups) from Cys122, Cys124 and Cys132, others residues such as Asp169, Asp120 and Ser168 are fundamental in the catalytic pocket composition, since they showed high attractive interaction energy with Zn2+ ion.
author2 91206359072
author_facet 91206359072
Barbosa, Emmanuel Duarte
author Barbosa, Emmanuel Duarte
author_sort Barbosa, Emmanuel Duarte
title Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
title_short Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
title_full Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
title_fullStr Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
title_full_unstemmed Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana
title_sort descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?on zn2+ na enzima alad humana
publisher PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA
publishDate 2017
url https://repositorio.ufrn.br/jspui/handle/123456789/21908
work_keys_str_mv AT barbosaemmanuelduarte descriobioqumicaqunticadobolsodeinteraodoonzn2naenzimaaladhumana
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spelling ndltd-IBICT-oai-repositorio.ufrn.br-123456789-219082018-05-23T23:28:43Z Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana Barbosa, Emmanuel Duarte 91206359072 Fulco, Umberto Laino 67196675487 Sinigaglia, Marialva 60901691020 Amaral, Viviane Souza do CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA ALAD Energia de liga??o MFCC DFT Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-02-02T13:30:50Z No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-02-08T19:26:36Z (GMT) No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) Made available in DSpace on 2017-02-08T19:26:36Z (GMT). No. of bitstreams: 1 EmmanuelDuarteBarbosa_DISSERT.pdf: 9706329 bytes, checksum: cf979f942793c968afbd04719854d7f0 (MD5) Previous issue date: 2016-07-29 A enzima Delta Aminolevul?nico Desidratase (ALAD) ? uma metaloprote?na citos?lica essencial em v?rios processos biol?gicos, uma vez que ? respons?vel pelo segundo passo da cat?lise enzim?tica na forma??o de porfobilinog?nio, um precursor dos tetrapirr?licos (heme, clorofila). Esta enzima ? bastante sens?vel a metais pesados e tem sido classicamente usada como um marcador na intoxica??o por chumbo. Sua inibi??o se d? pela substitui??o desses metais pesados no s?tio de liga??o a metais. Na ALAD humana, o Zinco (Zn2+) ocupa funcionalmente este s?tio sendo essencial para a coordena??o das cadeias de ?cido aminolevul?nico durante a cat?lise enzim?tica. Embora muitos ensaios in vitro, in vivo e in s?lico j? tenham demonstrado a import?ncia do Zn2+ nesse s?tio, n?o se tinha conhecimento de nenhum estudo baseado em abordagem qu?ntica com o intuito de elucidar esta intera??o de forma mais detalhada. Diante disso, o presente trabalho teve como objetivo analisar as muta??es missense que acometem o s?tio de liga??o ao zinco e descrever atrav?s de m?todos qu?nticos a energia de intera??o entre a enzima e o zinco com maior acur?cia utilizando o m?todo do Fracionamento Molecular com Capas Conjugadas (MFCC), quantificando energeticamente os res?duos de amino?cidos posicionados at? uma dist?ncia de 8,5 ? do centroide do ligante. Foi identificado as altera??es bioqu?micas na estrutura monom?rica dos mutantes, as quais resultam na diminui??o da atividade enzim?tica. Foram identificados um total de 30 res?duos com valores energ?ticos variados que interagem com o zinco no bols?o de liga??o. Aqueles que apresentaram valores significativos (de atra??o ou repuls?o) e est?o relacionados funcionalmente ? atividade enzim?tica foram: Lis199, Lis252, Arg 209, Arg 174, Cis122, Cis124 e Cis132; e aqueles que demonstraram relev?ncia para a perman?ncia do ?on no s?tio de liga??o foram: Asp169, Gli130, Gli133, Asp120 e Ser168. A partir disso, p?de-se concluir que al?m dos grupos nucle?filos (grupos tiolatos) dos res?duos Cis122, Cis124 e Cis132, os res?duos Asp169, Asp120 e Ser168 s?o fundamentais na composi??o do bols?o, uma vez que demonstraram grande quantidade de energia de intera??o atrativa com o ?on Zn2+. The enzyme Delta Aminolevulinic Dehydratase (ALAD) is a cytosolic metalloproteinase essential in several biological processes since it participates in the second step in porphobilinogen formation pathway, a tetrapyrrolic precursor of heme and chlorophyll. This enzyme is very sensitive to heavy metals and has traditionally been used as a biomarker in lead poisoning. Its inhibition occurs when these heavy metals are replaced inside the metal binding site. In human ALAD, Zinc (Zn2+) functionally occupies this site and it is essential for coordination of two chains of aminolevulinic acid for the enzymatic catalysis. Although many in vitro, in vivo and in silico works have already demonstrated the importance of Zn2+ at that site, to the best of our knowledge, there isn?t any studies on literature based on quantum approach in order to elucidate this interactions in more details. Therefore, the aim of the present study was to analyse the missense mutations that affect the zinc binding site and describe through quantum methods the energy interaction between zinc and ALAD with greater accuracy using the method of Molecular fractionation with conjugated caps (MFCC) by quantifying amino acid residues? energy positioned at 8.5 ? of distance with the ligand centroid. It was identified biochemical changes in the monomeric structure of mutants, which result in decreased enzyme activity. It were identified a total of 30 residues with a wide range of energy values. The residues with significant (atractition or repulsion) values and functionally related to enzymatic activity were: Lys199, Lys252, Cys122, Cys124 and Cys132; and those that demonstrated relevance to the ion permanence inside the binding site were: Asp169, Gly130, Gly133, Asp120 and Ser168. Thus, it could be concluded that in addition to the nucleophilic groups (thiolates groups) from Cys122, Cys124 and Cys132, others residues such as Asp169, Asp120 and Ser168 are fundamental in the catalytic pocket composition, since they showed high attractive interaction energy with Zn2+ ion. 2017-02-08T19:26:36Z 2017-02-08T19:26:36Z 2016-07-29 info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/masterThesis BARBOSA, Emmanuel Duarte. Descri??o bioqu?mica qu?ntica do bols?o de intera??o do ?ON Zn2+ na enzima ALAD humana. 2016. 51f. Disserta??o (Mestrado em Bioqu?mica) - Centro de Bioci?ncias, Universidade Federal do Rio Grande do Norte, Natal, 2016. https://repositorio.ufrn.br/jspui/handle/123456789/21908 por info:eu-repo/semantics/openAccess PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA UFRN Brasil reponame:Repositório Institucional da UFRN instname:Universidade Federal do Rio Grande do Norte instacron:UFRN