title: Crystal structure of SgcJ, an NTF2-like superfamily protein involved in biosynthesis of the nine-membered enediyne antitumor antibiotic C-1027.
abstract: Comparative analysis of the enediyne biosynthetic gene clusters revealed sets of conserved genes serving as outstanding candidates for the enediyne core. Here we report the crystal structures of SgcJ and its homologue NCS-Orf16, together with gene inactivation and site-directed mutagenesis studies, to gain insight into enediyne core biosynthesis. Gene inactivation in vivo establishes that SgcJ is required for C-1027 production in Streptomyces globisporus. SgcJ and NCS-Orf16 share a common structure with the nuclear transport factor 2-like superfamily of proteins, featuring a putative substrate binding or catalytic active site. Site-directed mutagenesis of the conserved residues lining this site allowed us to propose that SgcJ and its homologues may play a catalytic role in transforming the linear polyene intermediate, along with other enediyne polyketide synthase-associated enzymes, into an enzyme-sequestered enediyne core intermediate. These findings will help formulate hypotheses and design experiments to ascertain the function of SgcJ and its homologues in nine-membered enediyne core biosynthesis.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Tingting
authors.initials: T
authors.lastname: Huang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Chin-Yuan
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Jeremy R
authors.initials: JR
authors.lastname: Lohman
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Rudolf
authors.affiliation: Center for Structural Genomics of Infectious Diseases, University of Chicago, Chicago, IL USA.|Structural Biology Center, Argonne National Laboratory, Argonne, IL, USA.|Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Youngchang
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Structural Biology Center, Argonne National Laboratory, Argonne, IL, USA.|Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Changsoo
authors.initials: C
authors.lastname: Chang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Dong
authors.initials: D
authors.lastname: Yang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Ma
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Yan
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Ivana
authors.initials: I
authors.lastname: Crnovcic
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Lance
authors.initials: L
authors.lastname: Bigelow
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Shonda
authors.initials: S
authors.lastname: Clancy
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Craig A
authors.initials: CA
authors.lastname: Bingman
authors.affiliation: BioSciences at Rice and Department of Chemistry, Rice University, Houston, TX, USA.
authors.forename: Ragothaman M
authors.initials: RM
authors.lastname: Yennamalli
authors.affiliation: Center for Structural Genomics of Infectious Diseases, University of Chicago, Chicago, IL USA.|Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Gyorgy
authors.initials: G
authors.lastname: Babnigg
authors.affiliation: Center for Structural Genomics of Infectious Diseases, University of Chicago, Chicago, IL USA.|Structural Biology Center, Argonne National Laboratory, Argonne, IL, USA.|Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, Argonne, IL, USA.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Joachimiak
authors.affiliation: BioSciences at Rice and Department of Chemistry, Rice University, Houston, TX, USA.
authors.forename: George N
authors.initials: GN
authors.lastname: Phillips
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, FL, USA.|Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, FL, USA.|Natural Products Library Initiative at The Scripps Research Institute, The Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Ben
authors.initials: B
authors.lastname: Shen
mesh_terms: D000595:Amino Acid Sequence; D000617:Aminoglycosides / Q000096:biosynthesis*; D000903:Antibiotics, Antineoplastic / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D053281:Enediynes; D011090:Polyenes / Q000737:chemistry; D048630:Polyketide Synthases / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
chemical_list: D000617:Aminoglycosides; D000903:Antibiotics, Antineoplastic; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D053281:Enediynes; D011090:Polyenes; C058024:C 1027; D048630:Polyketide Synthases