{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T06:16:13Z","timestamp":1788416173669,"version":"build-2803163510"},"reference-count":41,"publisher":"American Chemical Society (ACS)","issue":"26","license":[{"start":{"date-parts":[[2015,6,19]],"date-time":"2015-06-19T00:00:00Z","timestamp":1434672000000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM035906"],"award-info":[{"award-number":["GM035906"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM106112"],"award-info":[{"award-number":["GM106112"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"publisher","award":["F-1511"],"award-info":[{"award-number":["F-1511"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"publisher","award":["F-1712"],"award-info":[{"award-number":["F-1712"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,7,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Spinosyns A and D (spinosad) are complex polyketide natural products biosynthesized through the cooperation of a modular polyketide synthase and several tailoring enzymes. SpnP catalyzes the final tailoring step, transferring forosamine from a TDP-d-forosamine donor substrate to a spinosyn pseudoaglycone acceptor substrate. Sequence analysis indicated that SpnP belongs to a small group of glycosyltransferases (GTs) that require an auxiliary protein for activation. However, unlike other GTs in this subgroup, no putative auxiliary protein gene could be located in the biosynthetic gene cluster. To learn more about SpnP, the structures of SpnP and its complex with TDP were determined to 2.50 and 3.15 \u00c5 resolution, respectively. Binding of TDP causes the reordering of several residues in the donor substrate pocket. SpnP possesses a structural feature that has only been previously observed in the related glycosyltransferase EryCIII, in which it mediates association with the auxiliary protein EryCII. This motif, H-X-R-X5-D-X5-R-X12\u201320-D-P-X3-W-L-X12\u201318-E-X4-G, may be predictive of glycosyltransferases that interact with an auxiliary protein. A reverse glycosyl transfer assay demonstrated that SpnP possesses measurable activity in the absence of an auxiliary protein. Our data suggest that SpnP can bind its donor substrate by itself but that the glycosyl transfer reaction is facilitated by an auxiliary protein that aids in the correct folding of a flexible loop surrounding the pseudoaglycone acceptor substrate-binding pocket.<\/jats:p>","DOI":"10.1021\/bi5003629","type":"journal-article","created":{"date-parts":[[2014,6,19]],"date-time":"2014-06-19T10:36:47Z","timestamp":1403174207000},"page":"4292-4301","source":"Crossref","is-referenced-by-count":26,"title":["Structural Studies of the Spinosyn Forosaminyltransferase,\nSpnP"],"prefix":"10.1021","volume":"53","author":[{"given":"Eta A.","family":"Isiorho","sequence":"first","affiliation":[{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]},{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Byung-Sun","family":"Jeon","sequence":"additional","affiliation":[{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]},{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nam Ho","family":"Kim","sequence":"additional","affiliation":[{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hung-wen","family":"Liu","sequence":"additional","affiliation":[{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]},{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian T.","family":"Keatinge-Clay","sequence":"additional","affiliation":[{"name":"The University of Texas at Austin , , , ,","place":["Austin, Texas, United States, 78712"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,6,19]]},"reference":[{"key":"2026090301462529900_cit1","doi-asserted-by":"crossref","first-page":"4839","DOI":"10.1016\/S0040-4039(00)93474-9","article-title":"A83543A-D,\nunique fermentation-derived tetracyclic macrolides","volume":"32","author":"Kirst","year":"1991","journal-title":"Tetrahedron Lett."},{"key":"2026090301462529900_cit2","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1002\/ps.358","article-title":"Natural products as insecticides:\nThe biology, biochemistry and quantitative structure-activity relationships\nof spinosyns and spinosoids","volume":"57","author":"Sparks","year":"2001","journal-title":"Pest Manage. Sci."},{"key":"2026090301462529900_cit3","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1023\/A:1010289901631","article-title":"A cluster of genes for the biosynthesis\nof spinosyns, novel macrolide insect control agents produced by Saccharopolyspora spinosa","volume":"78","author":"Waldron","year":"2000","journal-title":"Antonie van Leeuwenhoek"},{"key":"2026090301462529900_cit4","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/S1074-5521(01)00029-1","article-title":"Cloning and analysis of the spinosad biosynthetic gene cluster of Saccharopolyspora spinosa","volume":"8","author":"Waldron","year":"2001","journal-title":"Chem. Biol."},{"key":"2026090301462529900_cit5","doi-asserted-by":"crossref","first-page":"14582","DOI":"10.1021\/ja076580i","article-title":"The biosynthesis\nof spinosyn in Saccharopolyspora spinosa: Synthesis\nof the cross-bridging precursor and identification of the function\nof SpnJ","volume":"129","author":"Kim","year":"2007","journal-title":"J. Am. Chem. Soc."},{"key":"2026090301462529900_cit6","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1038\/nature09981","article-title":"Enzyme-catalysed\n[4+2] cycloaddition is a key step in the biosynthesis of spinosyn\nA","volume":"473","author":"Kim","year":"2011","journal-title":"Nature"},{"key":"2026090301462529900_cit7","doi-asserted-by":"crossref","first-page":"171","DOI":"10.7164\/antibiotics.53.171","article-title":"Synthesis and insecticidal\nactivity of spinosyn analogs functionally altered at the 2\u2032-,\n3\u2032- and 4\u2032-positions of the rhamnose moiety","volume":"53","author":"Creemer","year":"2000","journal-title":"J. Antibiot."},{"key":"2026090301462529900_cit8","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1093\/jee\/91.6.1277","article-title":"Biological activity of the spinosyns,\nnew fermentation derived insect control agents, on tobacco budworm\n(Lepidopter: Noctuidae) larvae","volume":"91","author":"Sparks","year":"1998","journal-title":"J. Econ. Entomol."},{"key":"2026090301462529900_cit9","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1146\/annurev.biochem.76.061005.092322","article-title":"Glycosyltransferases:\nStructures, functions, and mechanisms","volume":"77","author":"Lairson","year":"2008","journal-title":"Annu.\nRev. Biochem."},{"key":"2026090301462529900_cit10","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S0022-2836(03)00307-3","article-title":"An evolving\nhierarchical family classification for glycosyltransferases","volume":"328","author":"Coutinho","year":"2003","journal-title":"J. Mol. Biol."},{"key":"2026090301462529900_cit11","doi-asserted-by":"crossref","first-page":"D233","DOI":"10.1093\/nar\/gkn663","article-title":"The Carbohydrate-Active EnZymes database (CAZy): An\nexpert resource for glycogenomics","volume":"37","author":"Cantarel","year":"2009","journal-title":"Nucleic Acids\nRes."},{"key":"2026090301462529900_cit12","doi-asserted-by":"crossref","first-page":"7352","DOI":"10.1074\/jbc.M808441200","article-title":"Functional\ncharacterization and substrate specificity of spinosyn rhamnosyltransferase\nby in vitro reconstitution of spinosyn biosynthetic\nenzymes","volume":"284","author":"Chen","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2026090301462529900_cit13","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1021\/bi201860q","article-title":"Structural\nstudies of the spinosyn\nrhamnosyltransferase, SpnG","volume":"51","author":"Isiorho","year":"2012","journal-title":"Biochemistry"},{"key":"2026090301462529900_cit14","doi-asserted-by":"crossref","first-page":"17649","DOI":"10.1073\/pnas.1108484108","article-title":"Complete set\nof glycosyltransferase structures in the calicheamicin biosynthetic\npathway reveals the origin of regiospecificity","volume":"108","author":"Chang","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090301462529900_cit15","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.jmb.2011.10.036","article-title":"Structure of the glycosyltransferase\nEryCIII in complex with its activating P450 homologue EryCII","volume":"415","author":"Moncrieffe","year":"2012","journal-title":"J. Mol. Biol."},{"key":"2026090301462529900_cit16","doi-asserted-by":"crossref","first-page":"5336","DOI":"10.1073\/pnas.0607897104","article-title":"The crystal structure of two macrolide\nglycosyltransferases provides a blueprint for host cell antibiotic\nimmunity","volume":"204","author":"Bolam","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090301462529900_cit17","doi-asserted-by":"crossref","first-page":"6534","DOI":"10.1021\/ja049967j","article-title":"Characterization\nof the glycosyltransferase activity of DesVII: Analysis of and implication\nfor the biosynthesis of macrolide antibiotics","volume":"126","author":"Borisova","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026090301462529900_cit18","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/S0378-1119(96)00595-1","article-title":"Analysis of four tylosin biosynthetic\ngenes from the tylLM region of the Streptomyces fradiae genome","volume":"184","author":"Gandecha","year":"1997","journal-title":"Gene"},{"key":"2026090301462529900_cit19","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.jmb.2007.06.005","article-title":"Structure and action of the C-C bond-forming\nglycosyltransferase UrdGT2 involved in the biosynthesis of the antibiotic\nurdamycin","volume":"372","author":"Mittler","year":"2007","journal-title":"J. Mol. Biol."},{"key":"2026090301462529900_cit20","doi-asserted-by":"crossref","first-page":"9238","DOI":"10.1073\/pnas.1233577100","article-title":"Structure of the TDP-epi-vancosaminyltransferase\nGtfA\nfrom the chloroeremomycin biosynthetic pathway","volume":"100","author":"Mulichak","year":"2003","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090301462529900_cit21","doi-asserted-by":"crossref","first-page":"4954","DOI":"10.1021\/ja0771383","article-title":"In vivo characterization of the enzymes\ninvolved in TDP-d-forosamine biosynthesis in the spinosyn\npathway of Saccharopolyspora spinosa","volume":"130","author":"Hong","year":"2008","journal-title":"J. Am. Chem. Soc."},{"key":"2026090301462529900_cit22","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1016\/j.chembiol.2005.02.016","article-title":"AknT is an activating protein for\nthe glycosyltransferase AknS in l-aminodeoxysugar transfer\nto the aglycone of aclacinomycin A","volume":"12","author":"Lu","year":"2005","journal-title":"Chem. Biol."},{"key":"2026090301462529900_cit23","doi-asserted-by":"crossref","first-page":"2748","DOI":"10.1002\/anie.200503195","article-title":"Substrate\nspecificity of the macrolide-glycosylating\nenzyme pair DesVII\/DesVIII: Opportunities, limitations, and mechanistic\nhypotheses","volume":"45","author":"Borisova","year":"2006","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026090301462529900_cit24","doi-asserted-by":"crossref","first-page":"8071","DOI":"10.1021\/bi1007657","article-title":"Characterization\nof glycosyltransferase DesVII and\nits auxiliary partner protein DesVIII in the methymycin\/picromycin\nbiosynthetic pathway","volume":"49","author":"Borisova","year":"2010","journal-title":"Biochemistry"},{"key":"2026090301462529900_cit25","doi-asserted-by":"crossref","first-page":"14128","DOI":"10.1021\/ja053704n","article-title":"In vitro reconstitution of EryCIII\nactivity for the preparation of unnatural macrolides","volume":"127","author":"Yuan","year":"2005","journal-title":"J. Am. Chem. Soc."},{"key":"2026090301462529900_cit26","doi-asserted-by":"crossref","first-page":"16726","DOI":"10.1021\/ja043900e","article-title":"Characterization of tylM3\/tylM2 and\nmydC\/mycB pairs required for efficient glycosyltransfer in macrolide\nantibiotic biosynthesis","volume":"126","author":"Melanc\u0327on","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026090301462529900_cit27","doi-asserted-by":"crossref","first-page":"795","DOI":"10.7164\/antibiotics.51.795","article-title":"Conversion\nof spinosyn A and spinosyn\nD to their respective 9- and 17-pseudoaglycones and their aglycones","volume":"51","author":"Creemer","year":"1998","journal-title":"J. Antibiot."},{"key":"2026090301462529900_cit28","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S0076-6879(97)76066-X","article-title":"Processing of X-ray diffraction data collected in oscillation mode","volume":"276","author":"Otwinowski","year":"1997","journal-title":"Methods Enzymol."},{"key":"2026090301462529900_cit29","first-page":"1131","article-title":"A graphical user interface\nto the CCP4 program suite","volume":"D59","author":"Potterton","year":"2003","journal-title":"Acta Crystallogr."},{"key":"2026090301462529900_cit30","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1107\/S0021889807021206","article-title":"Phaser crystallographic\nsoftware","volume":"40","author":"McCoy","year":"2007","journal-title":"J. Appl.\nCrystallogr."},{"key":"2026090301462529900_cit31","first-page":"2222","article-title":"Towards complete\nvalidated models in the next generation of ARP\/wARP","volume":"D60","author":"Cohen","year":"2004","journal-title":"Acta Crystallogr."},{"key":"2026090301462529900_cit32","first-page":"2126","article-title":"Coot: Model-building tools for molecular graphics","volume":"D60","author":"Emsley","year":"2004","journal-title":"Acta Crystallogr."},{"key":"2026090301462529900_cit33","first-page":"2184","article-title":"REFMAC5 dictionary:\nOrganization of prior chemical knowledge and guidelines for its use","volume":"D60","author":"Vagin","year":"2004","journal-title":"Acta Crystallogr."},{"key":"2026090301462529900_cit34","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1002\/prot.24289","article-title":"Crystal structure of SsfS6, the putative C-glycosyltransferase involved\nin SF2575 biosynthesis","volume":"81","author":"Wang","year":"2013","journal-title":"Proteins"},{"key":"2026090301462529900_cit35","doi-asserted-by":"crossref","first-page":"3251","DOI":"10.1111\/j.1742-4658.2012.08711.x","article-title":"Crystal structure of the glycosyltransferase\nSnogD\nfrom the biosynthetic pathway of nogalamycin in Streptomyces nogalater","volume":"279","author":"Claesson","year":"2012","journal-title":"FEBS J."},{"key":"2026090301462529900_cit36","doi-asserted-by":"crossref","first-page":"5170","DOI":"10.1021\/bi036130c","article-title":"Crystal structure of\nvancosaminyltransferase GtfD from the vancomycin biosynthetic pathway:\nInteractions with acceptor and nucleotide ligands","volume":"43","author":"Mulichak","year":"2004","journal-title":"Biochemistry"},{"key":"2026090301462529900_cit37","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1038\/sj.emboj.7600970","article-title":"Structure\nof a flavonoid glucosyltransferase reveals the basis for plant natural\nproduct modification","volume":"25","author":"Offen","year":"2006","journal-title":"EMBO J."},{"key":"2026090301462529900_cit38","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1016\/S0969-2126(01)00616-5","article-title":"Structure of the UDP-glucosyltransferase\nGtfB that modifies the heptapeptide aglycone in the biosynthesis of\nvancomycin group antibiotics","volume":"9","author":"Mulichak","year":"2001","journal-title":"Structure"},{"key":"2026090301462529900_cit39","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1126\/science.1130028","article-title":"Exploiting the reversibility\nof natural product glycosyltransferase-catalyzed reactions","volume":"313","author":"Zhang","year":"2006","journal-title":"Science"},{"key":"2026090301462529900_cit40","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local\nalignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"2026090301462529900_cit41","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.gene.2006.08.021","article-title":"Functional analysis\nof desVIII homologues involved in glycosylation of macrolide antibiotics\nby interspecies complementation","volume":"386","author":"Hong","year":"2007","journal-title":"Gene"}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/bi5003629","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/53\/26\/4292\/15693983\/bi5003629.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/53\/26\/4292\/15693983\/bi5003629.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:46:38Z","timestamp":1788414398000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/53\/26\/4292\/1476814\/Structural-Studies-of-the-Spinosyn"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,6,19]]},"references-count":41,"journal-issue":{"issue":"26","published-online":{"date-parts":[[2014,6,19]]},"published-print":{"date-parts":[[2014,7,8]]}},"URL":"https:\/\/doi.org\/10.1021\/bi5003629","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,6,19]]}}}