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Dirigent isoflavene-forming PsPTS2: 3D structure, stereochemical, and kinetic characterization comparison with pterocarpan-forming PsPTS1 homolog in pea.
Meng, Qingyan; Moinuddin, Syed G A; Celoy, Rhodesia M; Smith, Clyde A; Young, Robert P; Costa, Michael A; Freeman, Rachel A; Fukaya, Masashi; Kim, Doo Nam; Cort, John R; Hawes, Martha C; van Etten, Hans D; Pandey, Pankaj; Chittiboyina, Amar G; Ferreira, Daneel; Davin, Laurence B; Lewis, Norman G.
Afiliación
  • Meng Q; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Moinuddin SGA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Celoy RM; School of Plant Sciences, University of Arizona, Tucson, Arizona, USA.
  • Smith CA; Stanford Synchrotron Radiation Lightsource, Stanford University, Menlo Park, California, USA.
  • Young RP; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington, USA.
  • Costa MA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Freeman RA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Fukaya M; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Kim DN; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington, USA.
  • Cort JR; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington, USA.
  • Hawes MC; School of Plant Sciences, University of Arizona, Tucson, Arizona, USA.
  • van Etten HD; School of Plant Sciences, University of Arizona, Tucson, Arizona, USA.
  • Pandey P; National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, Mississippi, USA.
  • Chittiboyina AG; National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, Mississippi, USA.
  • Ferreira D; National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, Mississippi, USA; Division of Pharmacognosy, Department of BioMolecular Sciences, School of Pharmacy, University of Mississippi, University, Mississippi, USA.
  • Davin LB; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
  • Lewis NG; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA. Electronic address: lewisn@wsu.edu.
J Biol Chem ; 300(3): 105647, 2024 Mar.
Article en En | MEDLINE | ID: mdl-38219818
ABSTRACT
Pea phytoalexins (-)-maackiain and (+)-pisatin have opposite C6a/C11a configurations, but biosynthetically how this occurs is unknown. Pea dirigent-protein (DP) PsPTS2 generates 7,2'-dihydroxy-4',5'-methylenedioxyisoflav-3-ene (DMDIF), and stereoselectivity toward four possible 7,2'-dihydroxy-4',5'-methylenedioxyisoflavan-4-ol (DMDI) stereoisomers was investigated. Stereoisomer configurations were determined using NMR spectroscopy, electronic circular dichroism, and molecular orbital analyses. PsPTS2 efficiently converted cis-(3R,4R)-DMDI into DMDIF 20-fold faster than the trans-(3R,4S)-isomer. The 4R-configured substrate's near ß-axial OH orientation significantly enhanced its leaving group abilities in generating A-ring mono-quinone methide (QM), whereas 4S-isomer's α-equatorial-OH was a poorer leaving group. Docking simulations indicated that the 4R-configured ß-axial OH was closest to Asp51, whereas 4S-isomer's α-equatorial OH was further away. Neither cis-(3S,4S)- nor trans-(3S,4R)-DMDIs were substrates, even with the former having C3/C4 stereochemistry as in (+)-pisatin. PsPTS2 used cis-(3R,4R)-7,2'-dihydroxy-4'-methoxyisoflavan-4-ol [cis-(3R,4R)-DMI] and C3/C4 stereoisomers to give 2',7-dihydroxy-4'-methoxyisoflav-3-ene (DMIF). DP homologs may exist in licorice (Glycyrrhiza pallidiflora) and tree legume Bolusanthus speciosus, as DMIF occurs in both species. PsPTS1 utilized cis-(3R,4R)-DMDI to give (-)-maackiain 2200-fold more efficiently than with cis-(3R,4R)-DMI to give (-)-medicarpin. PsPTS1 also slowly converted trans-(3S,4R)-DMDI into (+)-maackiain, reflecting the better 4R configured OH leaving group. PsPTS2 and PsPTS1 provisionally provide the means to enable differing C6a and C11a configurations in (+)-pisatin and (-)-maackiain, via identical DP-engendered mono-QM bound intermediate generation, which PsPTS2 either re-aromatizes to give DMDIF or PsPTS1 intramolecularly cyclizes to afford (-)-maackiain. Substrate docking simulations using PsPTS2 and PsPTS1 indicate cis-(3R,4R)-DMDI binds in the anti-configuration in PsPTS2 to afford DMDIF, and the syn-configuration in PsPTS1 to give maackiain.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas de Plantas / Pisum sativum / Pterocarpanos Idioma: En Revista: J Biol Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas de Plantas / Pisum sativum / Pterocarpanos Idioma: En Revista: J Biol Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos