An Unusual Role for a Mobile Flavin in StaC-like Indolocarbazole Biosynthetic Enzymes
Name
Drennan_An unusual role.pdf
Size
1.81 MB
Format
Adobe PDF
Checksum (MD5)
a2ac2b2ba174ce23e1858816a460c603
Author(s)
Goldman, Peter J.
Hamill, Michael J.
Howard-Jones, Annaleise R.
Walsh, Christopher T.
Elliott, Sean J.
Drennan, Catherine L.
Ryan, Katherine S.
Ryan, Katherine S.
Date Issued
July 2012
Journal
Chemistry & Biology
Publisher
Springer Science + Business Media B.V.
Citation
Goldman, Peter J., Katherine S. Ryan, Michael J. Hamill, Annaleise R. Howard-Jones, Christopher T. Walsh, Sean J. Elliott, and Catherine L. Drennan. “An Unusual Role for a Mobile Flavin in StaC-like Indolocarbazole Biosynthetic Enzymes.” Chemistry & Biology 19, no. 7 (July 2012): 855-865.
Version
Author's final manuscript
Abstract
The indolocarbazole biosynthetic enzymes StaC, InkE, RebC, and AtmC mediate the degree of oxidation of chromopyrrolic acid on route to the natural products staurosporine, K252a, rebeccamycin, and AT2433-A1, respectively. Here, we show that StaC and InkE, which mediate a net 4-electron oxidation, bind FAD with a micromolar Kd, whereas RebC and AtmC, which mediate a net 8-electron oxidation, bind FAD with a nanomolar Kd while displaying the same FAD redox properties. We further create RebC-10x, a RebC protein with ten StaC-like amino acid substitutions outside of previously characterized FAD-binding motifs and the complementary StaC-10x. We find that these mutations mediate both FAD affinity and product specificity, with RebC-10x displaying higher StaC activity than StaC itself. X-ray structures of this StaC catalyst identify the substrate of StaC as 7-carboxy-K252c and suggest a unique mechanism for this FAD-dependent enzyme.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/j.chembiol.2012.05.016
