{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T15:30:19Z","timestamp":1787844619513,"version":"build-2784847793"},"reference-count":23,"publisher":"Oxford University Press (OUP)","issue":"9","license":[{"start":{"date-parts":[[2009,5,15]],"date-time":"2009-05-15T00:00:00Z","timestamp":1242345600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2009,5,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The biogenesis of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) is accomplished by the methylerythritol phosphate (MEP) pathway in plants, bacteria and parasites, making it a potential target for the development of anti-infective agents and herbicides. The biosynthetic enzymes comprising this pathway catalyze intriguing chemical transformations on diphosphate scaffolds, offering an opportunity to generate novel analogs in this synthetically challenging compound class. Such a biosynthetic approach to generating new diphosphate analogs may involve transformation through discrete diphosphate species, presenting unique challenges in structure determination and characterization of unnatural enzyme-generated diphosphate products produced in tandem. We have developed 1H\u201331P\u201331P correlation NMR spectroscopy techniques for the direct characterization of crude MEP pathway enzyme products at low concentrations (200 \u03bcM to 5 mM) on a room temperature (non-cryogenic) NMRprobe. Coupling the 100% natural abundance of the 31Pnucleus with the high intrinsic sensitivity of proton NMR, 1H\u201331P\u201331Pcorrelation spectroscopy is particularly useful for characterization of unnatural diphosphateenzyme products in the MEP pathway. As proof of principle, we demonstrate the rapid characterization of natural enzyme products of the enzymes IspD, E and F in tandem enzyme incubations. In addition, we have characterized several unnatural enzyme products using this technique, including new products of cytidyltransferase IspD bearing erythritol, glycerol and ribose components. The results of this study indicate that IspD may be a useful biocatalyst and highlight 1H\u201331P\u201331Pcorrelation spectroscopy as a valuable tool for the characterization of other unnatural products in non-mammalian isoprenoid biosynthesis.<\/jats:p>\n                  <jats:p\/>","DOI":"10.1039\/b903513c","type":"journal-article","created":{"date-parts":[[2009,5,15]],"date-time":"2009-05-15T08:30:16Z","timestamp":1242376216000},"page":"935-944","source":"Crossref","is-referenced-by-count":10,"title":["Probing phosphorylation by non-mammalian isoprenoid biosynthetic enzymes using 1H\u201331P\u201331P correlation NMR spectroscopy"],"prefix":"10.1093","volume":"5","author":[{"given":"Ananya","family":"Majumdar","sequence":"first","affiliation":[{"name":"Biomolecular NMR Center, The Johns Hopkins University a , Baltimore, MD 21218 USA \u00a0 ananya@jhu.edu"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meha H","family":"Shah","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine b , Baltimore, MD 21205 USA \u00a0 cmeyers@jhmi.edu"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Kipchirchir","family":"Bitok","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine b , Baltimore, MD 21205 USA \u00a0 cmeyers@jhmi.edu"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria E","family":"Hassis-LeBeau","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine b , Baltimore, MD 21205 USA \u00a0 cmeyers@jhmi.edu"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caren L","family":"Freel Meyers","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine b , Baltimore, MD 21205 USA \u00a0 cmeyers@jhmi.edu"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2009,5,15]]},"reference":[{"issue":"12","key":"2026012914023662600_cit1","doi-asserted-by":"crossref","first-page":"1401","DOI":"10.1007\/s00018-004-3381-z","article-title":"Biosynthesis of isoprenoids via the non-mevalonate pathway","volume":"61","author":"Eisenreich","year":"2004","journal-title":"Cell. 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