A Bowman-Birk protease inhibitor purified, cloned, sequenced and characterized from the seeds of Maclura pomifera (Raf.) Schneid

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dc.citation.issue2
dc.citation.volume245
dc.contributor.authorIndarte, Martin
dc.contributor.authorLazza, Cristian M.
dc.contributor.authorAssis, Diego [UNIFESP]
dc.contributor.authorCaffini, Nestor O.
dc.contributor.authorJuliano, Maria A. [UNIFESP]
dc.contributor.authorAviles, Francesc X.
dc.contributor.authorDaura, Xavier
dc.contributor.authorLopez, Laura M. I.
dc.contributor.authorTrejo, Sebastian A.
dc.coverageNew York
dc.date.accessioned2020-07-17T14:03:10Z
dc.date.available2020-07-17T14:03:10Z
dc.date.issued2017
dc.description.abstractA new BBI-type protease inhibitor with remarkable structural characteristics was purified, cloned, and sequenced from seeds of Maclura pomifera , a dicotyledonous plant belonging to the Moraceae family. In this work, we report a Bowman-Birk inhibitor (BBI) isolated, purified, cloned, and characterized from Maclura pomifera seeds (MpBBI), the first of this type from a species belonging to Moraceae family. MpBBI was purified to homogeneity by RP-HPLC, total RNA was extracted from seeds of M. pomifera, and the 3'RACE-PCR method was applied to obtain the cDNA, which was cloned and sequenced. Peptide mass fingerprinting (PMF) analysis showed correspondence between the in silico-translated protein and MpBBI, confirming that it corresponds to a new plant protease inhibitor. The obtained cDNA encoded a polypeptide of 65 residues and possesses 10 cysteine residues, with molecular mass of 7379.27, pI 6.10, and extinction molar coefficient of 9105 M-1 cm(-1). MpBBI inhibits strongly trypsin with K (i) in the 10(-10) M range and was stable in a wide array of pH and extreme temperatures. MpBBI comparative modeling was applied to gain insight into its 3D structure and highlighted some distinguishing features: (1) two non-identical loops, (2) loop 1 (CEEESRC) is completely different from any known BBI, and (3) the amount of disulphide bonds is also different from any reported BBI from dicot plants.en
dc.description.affiliationPHusisTherapeutics, 3210 Merryfield Row, San Diego, CA 92121 USA
dc.description.affiliationUniv Nacl La Plata, Fac Ciencias Exactas, Ctr Invest Proteinas Vegetales, RA-1900 La Plata, Buenos Aires, Argentina
dc.description.affiliationUniv Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo, Brazil
dc.description.affiliationUniv Autonoma Barcelona, IBB, E-08193 Barcelona, Spain
dc.description.affiliationUniv Nacl Arturo Jauretche, Inst Ciencias Salud, 1888 Florencio Varela, Buenos Aires, DF, Argentina
dc.description.affiliationCITEC, Gonnet, B1897, Buenos Aires, DF, Argentina
dc.description.affiliationInst Multidisciplinar Biol Celular IMBICE, Lab Neurofisiol, B1906APO, Buenos Aires, DF, Argentina
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)
dc.description.sponsorshipCYTED
dc.description.sponsorshipCONICET
dc.description.sponsorshipIDFAPESP: 12/50191-4R
dc.description.sponsorshipIDCYTED: Red tematica PROMAL 210RT0398
dc.description.sponsorshipIDCONICET: PIP 0297
dc.format.extent343-353
dc.identifierhttp://dx.doi.org/10.1007/s00425-016-2611-6
dc.identifier.citationPlanta. New York, v. 245, n. 2, p. 343-353, 2017.
dc.identifier.doi10.1007/s00425-016-2611-6
dc.identifier.issn0032-0935
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/55201
dc.identifier.wosWOS:000393691500008
dc.language.isoeng
dc.publisherSpringer
dc.relation.ispartofPlanta
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectBBI-type protease inhibitoren
dc.subjectCloningen
dc.subjectHomology modelingen
dc.subjectLoopen
dc.subjectThree-dimensional structureen
dc.subjectTrypsin inhibitionen
dc.titleA Bowman-Birk protease inhibitor purified, cloned, sequenced and characterized from the seeds of Maclura pomifera (Raf.) Schneiden
dc.typeinfo:eu-repo/semantics/article
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