title: Formation of a Novel Macrocyclic Alkaloid from the Unnatural Farnesyl Diphosphate Analogue Anilinogeranyl Diphosphate by 5-Epi-Aristolochene Synthase.
abstract: As part of an effort to identify substrate analogs suitable for helping to resolve structural features important for terpene synthases, the inhibition of 5-epi-aristolochene biosynthesis from farnesyl diphosphate (FPP) by the tobacco 5-epi-aristolochene synthase incubated with anilinogeranyl diphosphate (AGPP) was examined. The apparent noncompetitive nature of the inhibition supported further assessment of how AGPP might be bound to crystallographic forms of the enzyme. Surprisingly, the bound form of the inhibitor appeared to have undergone a cyclization event consistent with the native mechanism associated with FPP catalysis. Biocatalytic formation of a novel 13-membered macrocyclic paracyclophane alkaloid was confirmed by high-resolution GC-MS and NMR analysis. This work provides insights into new biosynthetic means for generating novel, functionally diversified, medium-sized terpene alkaloids.
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Rising
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Charisse M
authors.initials: CM
authors.lastname: Crenshaw
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Hyun Jo
authors.initials: HJ
authors.lastname: Koo
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Thangaiah
authors.initials: T
authors.lastname: Subramanian
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Kareem A H
authors.initials: KA
authors.lastname: Chehade
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Courtney
authors.initials: C
authors.lastname: Starks
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Keith D
authors.initials: KD
authors.lastname: Allen
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Andres
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: H Peter
authors.initials: HP
authors.lastname: Spielmann
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Noel
authors.affiliation: †Departments of Pharmaceutical Sciences, ‡Cellular and Molecular Biochemistry, §Chemistry, ⊥Center for Structural Biology, ∥Markey Cancer Center, University of Kentucky, Lexington, Lexington, Kentucky, United States.|∇Howard Hughes Medical Institute, Salk Institute, La Jolla, California 92037, United States.
authors.forename: Joe
authors.initials: J
authors.lastname: Chappell
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000378:metabolism*; D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000378:metabolism*; D047028:Macrocyclic Compounds / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism*; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism*; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism
chemical_list: D000470:Alkaloids; D047028:Macrocyclic Compounds; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D013729:Terpenes; C004808:farnesyl pyrophosphate; D019883:Alkyl and Aryl Transferases; C078217:5-epi-aristolochene synthase