Purification and biochemical properties of a Kunitz-type trypsin inhibitor from Entada acaciifolia (Benth.) seeds

dc.creatorRamalho de Oliveira, Caio Fernando
dc.creatorVasconcelos, Ilka Maria
dc.creatorAparicio, Ricardo
dc.creatorMachado Freire, Maria das Gracas
dc.creatorBaldasso, Paulo Aparecido
dc.creatorMarangoni, Sergio
dc.creatorRodrigues Macedo, Maria Ligia
dc.date2012
dc.date2013-09-19T18:06:35Z
dc.date2016-07-01T14:42:02Z
dc.date2013-09-19T18:06:35Z
dc.date2016-07-01T14:42:02Z
dc.date.accessioned2018-03-29T01:54:42Z
dc.date.available2018-03-29T01:54:42Z
dc.identifierProcess Biochemistry. Elsevier, v.47, n.6, p.929-935, 2012
dc.identifier1359-5113
dc.identifierWOS:000304511000005
dc.identifier10.1016/j.procbio.2012.02.022
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/2287
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/2287
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1308611
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionA new trypsin inhibitor (EATI) was isolated from Entada acaciifolia (Benth.) seeds. EATI is a competitive inhibitor with a molecular mass of 20 kDa and an inhibition stoichiometry of 1:1 for bovine trypsin. The dissociation constant (K-i) calculated was 1.75 nmol/L, displaying a high affinity between enzyme and inhibitor. Both Native PAGE and RP-HPLC revealed that EATI is composed of four isoinhibitors that share the amino acid composition and the amino-terminal sequence homolog to Kunitz-type inhibitors. EATI is stable to denaturation by heat (up to 70 degrees C), pH (2-10), urea (8 mol/L) and its inhibitory activity was unaltered in different concentrations of DTT (up to 100 mmol/L). CD analysis revealed that EATI in reduced form underwent structural modifications associated with a decrease in thermal and pH stabilities, suggesting that their disulfide bonds are not involved in the structuring of its reactive site, but are important for maintenance of its conformational stability. This behavior makes EATI one of the few inhibitors described in the literature with high DTT resistance. (C) 2012 Elsevier Ltd. All rights reserved.
dc.description47
dc.description6
dc.description929
dc.description935
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionFoundation to Support the Development of Education, Science and Technology of State of Mato Grosso do Sul (FUNDECT - Fundacao de Apoio ao Desenvolvimento do Ensino, Ciencia e Tecnologia do Estado de Mato Grosso do Sul)
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.languageeng
dc.publisherElsevier
dc.publisherOxford
dc.relationProcess Biochemistry
dc.rightsfechado
dc.sourceWOS
dc.subjectEntada acaciifolia
dc.subjectCircular dichroism
dc.subjectKunitz-type inhibitor
dc.subjectTrypsin inhibitor
dc.subjectAMINO-ACID-SEQUENCE
dc.subjectSINGLE DISULFIDE BRIDGE
dc.subjectPROTEINASE-INHIBITORS
dc.subjectPROTEASE INHIBITOR
dc.subjectCRYSTAL-STRUCTURE
dc.subjectLEPIDOPTERA-PYRALIDAE
dc.subjectSTABILITY
dc.subjectSERINE
dc.subjectSITE
dc.subjectFAMILY
dc.titlePurification and biochemical properties of a Kunitz-type trypsin inhibitor from Entada acaciifolia (Benth.) seeds
dc.typeArtículos de revistas


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