title: Biosynthetic potential of sesquiterpene synthases: product profiles of Egyptian Henbane premnaspirodiene synthase and related mutants.
abstract: The plant terpene synthase (TPS) family is responsible for the biosynthesis of a variety of terpenoid natural products possessing diverse biological functions. TPSs catalyze the ionization and, most commonly, rearrangement and cyclization of prenyl diphosphate substrates, forming linear and cyclic hydrocarbons. Moreover, a single TPS often produces several minor products in addition to a dominant product. We characterized the catalytic profiles of Hyoscyamus muticus premnaspirodiene synthase (HPS) and compared it with the profile of a closely related TPS, Nicotiana tabacum 5-epi-aristolochene synthase (TEAS). The profiles of two previously studied HPS and TEAS mutants, each containing nine interconverting mutations, dubbed HPS-M9 and TEAS-M9, were also characterized. All four TPSs were compared under varying temperature and pH conditions. In addition, we solved the X-ray crystal structures of TEAS and a TEAS quadruple mutant complexed with substrate and products to gain insight into the enzymatic features modulating product formation. These informative structures, along with product profiles, provide new insight into plant TPS catalytic promiscuity.
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Hyun Jo
authors.initials: HJ
authors.lastname: Koo
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.|Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Vickery
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Gordon V
authors.initials: GV
authors.lastname: Louie
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Paul E
authors.initials: PE
authors.lastname: O'Maille
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Marianne
authors.initials: M
authors.lastname: Bowman
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Charisse M
authors.initials: CM
authors.lastname: Nartey
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkart
authors.affiliation: Howard Hughes Medical Institute, The Salk Institute for Biological Studies, Jack H Skirball Center for Chemical Biology and Proteomics, La Jolla, CA, USA.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Noel
mesh_terms: D020134:Catalytic Domain; D004795:Enzyme Stability / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D032304:Hyoscyamus / Q000201:enzymology* / Q000235:genetics; D009154:Mutation; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012717:Sesquiterpenes / Q000378:metabolism*; D013696:Temperature
chemical_list: D010940:Plant Proteins; D012717:Sesquiterpenes