taxonID	type	format	identifier	references	title	description	created	creator	contributor	publisher	audience	source	license	rightsHolder	datasetID
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291740/files/figure.png	https://doi.org/10.5281/zenodo.8291740	Fig. 3. LC-MS analyses of in vitro assays with CYP72A proteins expressed in yeast microsomes. 100 μl reactions containing 10 μl CYP72A microsomes isolated from WAT11 with 250 μM loganic acid or loganin and 500 μM NADPH in 100 mM NaPO (pH 7.4) were incubated at 30◦C and analyzed by LC-MS as 4 described in the experimental procedures. (A) Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089). (B) Extracted ion chromatograms for loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246).	Fig. 3. LC-MS analyses of in vitro assays with CYP72A proteins expressed in yeast microsomes. 100 μl reactions containing 10 μl CYP72A microsomes isolated from WAT11 with 250 μM loganic acid or loganin and 500 μM NADPH in 100 mM NaPO (pH 7.4) were incubated at 30◦C and analyzed by LC-MS as 4 described in the experimental procedures. (A) Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089). (B) Extracted ion chromatograms for loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246).	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291736/files/figure.png	https://doi.org/10.5281/zenodo.8291736	Fig. 1. Proposed divergence of the TIA pathway between Camptotheca and Catharanthus. After 7-deoxyloganic acid hydroxylase (7DLH) converts 7-deoxyloganic acid to loganic acid, the pathways in these two species diverge. The Catharanthus pathway uses loganic acid methyltransferase (LAMT) to convert loganic acid into loganin and secologanin synthase (SLS) to convert loganin into secologanin. The Camptotheca pathway bypasses LAMT and uses secologanic acid synthase (SLAS) to metabolize loganic acid directly to secologanic acid.	Fig. 1. Proposed divergence of the TIA pathway between Camptotheca and Catharanthus. After 7-deoxyloganic acid hydroxylase (7DLH) converts 7-deoxyloganic acid to loganic acid, the pathways in these two species diverge. The Catharanthus pathway uses loganic acid methyltransferase (LAMT) to convert loganic acid into loganin and secologanin synthase (SLS) to convert loganin into secologanin. The Camptotheca pathway bypasses LAMT and uses secologanic acid synthase (SLAS) to metabolize loganic acid directly to secologanic acid.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291742/files/figure.png	https://doi.org/10.5281/zenodo.8291742	Fig. 4. LC-MS analyses of in vitro assays with purified His6-tagged CYP72A proteins reconstituted with His6-tagged CPR proteins. Reactions containing purified His6- tagged CYP72A protein, full-length His6-tagged Caa CPR1 protein (A, B) or full-length His6-tagged Caa CPR2 protein (C, D), and 250 μM loganic acid (A,C) or loganin (B,D), were incubated at 30◦C and analyzed by LC-MS as described in experimental procedures. Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089); loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246) are given with stacked chromatograms as marked.	Fig. 4. LC-MS analyses of in vitro assays with purified His6-tagged CYP72A proteins reconstituted with His6-tagged CPR proteins. Reactions containing purified His6- tagged CYP72A protein, full-length His6-tagged Caa CPR1 protein (A, B) or full-length His6-tagged Caa CPR2 protein (C, D), and 250 μM loganic acid (A,C) or loganin (B,D), were incubated at 30◦C and analyzed by LC-MS as described in experimental procedures. Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089); loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246) are given with stacked chromatograms as marked.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291744/files/figure.png	https://doi.org/10.5281/zenodo.8291744	Fig. 5. Area of loganic acid, loganin and products from in vitro reconstitution assays conducted with full-length Camptotheca His6-tagged CPR1. Integrated areas from LC-MS analyses of purified His6-tagged CYP72A proteins reconstituted with full-length His6-tagged Caa CPR1 are shown for no NADPH (gray) and plus NADPH (gray slashed) reactions.	Fig. 5. Area of loganic acid, loganin and products from in vitro reconstitution assays conducted with full-length Camptotheca His6-tagged CPR1. Integrated areas from LC-MS analyses of purified His6-tagged CYP72A proteins reconstituted with full-length His6-tagged Caa CPR1 are shown for no NADPH (gray) and plus NADPH (gray slashed) reactions.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291738/files/figure.png	https://doi.org/10.5281/zenodo.8291738	Fig. 2. CYP72A multiple sequence alignment. Signal anchor domain fusion of CYP72A565 into CYP72A564 is underlined; SRS regions are underlined in bold; predicted substrate contacts within 4.5 Å of loganic acid/loganin are gray-filled.	Fig. 2. CYP72A multiple sequence alignment. Signal anchor domain fusion of CYP72A565 into CYP72A564 is underlined; SRS regions are underlined in bold; predicted substrate contacts within 4.5 Å of loganic acid/loganin are gray-filled.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2E6BFE38B484.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291746/files/figure.png	https://doi.org/10.5281/zenodo.8291746	Fig. 6. Type I binding spectra of SLAS candidates. Substrate-induced Type I binding spectra for (A) CYP72A564, (B) CYP72A565 and from 998 nM (violet) to 1.90 mM (crimson) using loganic acid (top inset) or loganin (bottom inset). (C) Binding isotherms calculated as the difference of the valley (~419 nm) from the peak (~388 nm) are shown for loganic acid () and loganin (). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)	Fig. 6. Type I binding spectra of SLAS candidates. Substrate-induced Type I binding spectra for (A) CYP72A564, (B) CYP72A565 and from 998 nM (violet) to 1.90 mM (crimson) using loganic acid (top inset) or loganin (bottom inset). (C) Binding isotherms calculated as the difference of the valley (~419 nm) from the peak (~388 nm) are shown for loganic acid () and loganin (). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2C4AFABDB246.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291748/files/figure.png	https://doi.org/10.5281/zenodo.8291748	Fig. 7. Molecular models of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565. (A) Backbone overlays of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565 models are shown with the alpha-carbon RMSD amongst CYP72A1, CYP72A564 and CYP72A565 depicted from green (0.0 Å) to yellow (3.0 Å) to red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A564 (orange). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A565 (rose).	Fig. 7. Molecular models of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565. (A) Backbone overlays of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565 models are shown with the alpha-carbon RMSD amongst CYP72A1, CYP72A564 and CYP72A565 depicted from green (0.0 Å) to yellow (3.0 Å) to red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A564 (orange). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A565 (rose).	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2C4AFABDB246.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291750/files/figure.png	https://doi.org/10.5281/zenodo.8291750	Fig. 8. Molecular models of Camptotheca CYP72A564, CYP72A565 and CYP72A730. (A) Backbone overlays of Camptotheca CYP72A730, CYP72A564 and CYP72A565 models are shown with the RMSD variance of CYP72A564 and CYP72A565 from the CYP72A730 backbone depicted in green (0.0 Å), yellow (3.0 Å) and red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A564 (orange) versus loganic acid (aqua) docked in CYP72A730 (magenta). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A565 (rose) versus loganic acid (aqua) docked in CYP72A730 (magenta).	Fig. 8. Molecular models of Camptotheca CYP72A564, CYP72A565 and CYP72A730. (A) Backbone overlays of Camptotheca CYP72A730, CYP72A564 and CYP72A565 models are shown with the RMSD variance of CYP72A564 and CYP72A565 from the CYP72A730 backbone depicted in green (0.0 Å), yellow (3.0 Å) and red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A564 (orange) versus loganic acid (aqua) docked in CYP72A730 (magenta). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A565 (rose) versus loganic acid (aqua) docked in CYP72A730 (magenta).	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2C4AFABDB246.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291742/files/figure.png	https://doi.org/10.5281/zenodo.8291742	Fig. 4. LC-MS analyses of in vitro assays with purified His6-tagged CYP72A proteins reconstituted with His6-tagged CPR proteins. Reactions containing purified His6- tagged CYP72A protein, full-length His6-tagged Caa CPR1 protein (A, B) or full-length His6-tagged Caa CPR2 protein (C, D), and 250 μM loganic acid (A,C) or loganin (B,D), were incubated at 30◦C and analyzed by LC-MS as described in experimental procedures. Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089); loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246) are given with stacked chromatograms as marked.	Fig. 4. LC-MS analyses of in vitro assays with purified His6-tagged CYP72A proteins reconstituted with His6-tagged CPR proteins. Reactions containing purified His6- tagged CYP72A protein, full-length His6-tagged Caa CPR1 protein (A, B) or full-length His6-tagged Caa CPR2 protein (C, D), and 250 μM loganic acid (A,C) or loganin (B,D), were incubated at 30◦C and analyzed by LC-MS as described in experimental procedures. Extracted ion chromatograms for loganic acid (m/z 375.1297), secologanic acid (m/z 373.1140), secoxyloganic acid (m/z 389.1089); loganin sodium salt (m/z +413.1418), secologanin sodium salt (m/z +411.1262), secoxyloganin (m/z 403.1246) are given with stacked chromatograms as marked.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF671365FFCD2C4AFABDB246.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291744/files/figure.png	https://doi.org/10.5281/zenodo.8291744	Fig. 5. Area of loganic acid, loganin and products from in vitro reconstitution assays conducted with full-length Camptotheca His6-tagged CPR1. Integrated areas from LC-MS analyses of purified His6-tagged CYP72A proteins reconstituted with full-length His6-tagged Caa CPR1 are shown for no NADPH (gray) and plus NADPH (gray slashed) reactions.	Fig. 5. Area of loganic acid, loganin and products from in vitro reconstitution assays conducted with full-length Camptotheca His6-tagged CPR1. Integrated areas from LC-MS analyses of purified His6-tagged CYP72A proteins reconstituted with full-length His6-tagged Caa CPR1 are shown for no NADPH (gray) and plus NADPH (gray slashed) reactions.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF611361FFCD2944FF78B2C5.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291748/files/figure.png	https://doi.org/10.5281/zenodo.8291748	Fig. 7. Molecular models of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565. (A) Backbone overlays of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565 models are shown with the alpha-carbon RMSD amongst CYP72A1, CYP72A564 and CYP72A565 depicted from green (0.0 Å) to yellow (3.0 Å) to red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A564 (orange). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A565 (rose).	Fig. 7. Molecular models of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565. (A) Backbone overlays of Catharanthus CYP72A1 and Camptotheca CYP72A564 and CYP72A565 models are shown with the alpha-carbon RMSD amongst CYP72A1, CYP72A564 and CYP72A565 depicted from green (0.0 Å) to yellow (3.0 Å) to red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A564 (orange). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganin (aqua) docked in Catharanthus CYP72A1 (blue) and loganic acid (gray) docked in Camptotheca CYP72A565 (rose).	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF611361FFCD2944FF78B2C5.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291738/files/figure.png	https://doi.org/10.5281/zenodo.8291738	Fig. 2. CYP72A multiple sequence alignment. Signal anchor domain fusion of CYP72A565 into CYP72A564 is underlined; SRS regions are underlined in bold; predicted substrate contacts within 4.5 Å of loganic acid/loganin are gray-filled.	Fig. 2. CYP72A multiple sequence alignment. Signal anchor domain fusion of CYP72A565 into CYP72A564 is underlined; SRS regions are underlined in bold; predicted substrate contacts within 4.5 Å of loganic acid/loganin are gray-filled.	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
8D608791DF611361FFCD2944FF78B2C5.taxon	http://purl.org/dc/dcmitype/StillImage	image/png	https://zenodo.org/record/8291750/files/figure.png	https://doi.org/10.5281/zenodo.8291750	Fig. 8. Molecular models of Camptotheca CYP72A564, CYP72A565 and CYP72A730. (A) Backbone overlays of Camptotheca CYP72A730, CYP72A564 and CYP72A565 models are shown with the RMSD variance of CYP72A564 and CYP72A565 from the CYP72A730 backbone depicted in green (0.0 Å), yellow (3.0 Å) and red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A564 (orange) versus loganic acid (aqua) docked in CYP72A730 (magenta). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A565 (rose) versus loganic acid (aqua) docked in CYP72A730 (magenta).	Fig. 8. Molecular models of Camptotheca CYP72A564, CYP72A565 and CYP72A730. (A) Backbone overlays of Camptotheca CYP72A730, CYP72A564 and CYP72A565 models are shown with the RMSD variance of CYP72A564 and CYP72A565 from the CYP72A730 backbone depicted in green (0.0 Å), yellow (3.0 Å) and red (4.5 Å). (B) SRS regions in CYP72A proteins shown with predicted substrate contact residues (gray fill). (C) Identical versus (D) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A564 (orange) versus loganic acid (aqua) docked in CYP72A730 (magenta). (E) Identical versus (F) different side chain residues predicted within 4.5 Å of loganic acid (gray) docked in Camptotheca CYP72A565 (rose) versus loganic acid (aqua) docked in CYP72A730 (magenta).	2021-03-31	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.		Zenodo	biologists	Miller, Justin C.;Hollatz, Allison J.;Schuler, Mary A.			
