4EIP

Native and K252c bound RebC-10x

Summary for 4EIP

• Entry DOI: 10.2210/pdb4eip/pdb
• Related: 2R0C 2R0P 4EIQ
• Descriptor: Putative FAD-monooxygenase, 6,7,12,13-tetrahydro-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazol-5-one, FLAVIN-ADENINE DINUCLEOTIDE, ... (4 entities in total)
• Functional Keywords: flavin adenine dinucleotide, k252c, monooxygenase, indolocarbazole, oxidoreductase-oxidoreductase inhibitor complex, oxidoreductase/oxidoreductase inhibitor
• Biological source: Lechevalieria aerocolonigenes (Nocardia aerocolonigenes)
• Total number of polymer chains: 2
• Total formula weight: 121243.13

Authors

Goldman, P.J.,Ryan, K.S.,Howard-Jones, A.R.,Hamill, M.J.,Elliott, S.J.,Walsh, C.T.,Drennan, C.L. (deposition date: 2012-04-05, release date: 2012-08-08, Last modification date: 2024-02-28)

Primary citation

Goldman, P.J.,Ryan, K.S.,Hamill, M.J.,Howard-Jones, A.R.,Walsh, C.T.,Elliott, S.J.,Drennan, C.L.
An Unusual Role for a Mobile Flavin in StaC-like Indolocarbazole Biosynthetic Enzymes.
Chem.Biol., 19:855-865, 2012

PubMed 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 K(d), whereas RebC and AtmC, which mediate a net 8-electron oxidation, bind FAD with a nanomolar K(d) 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.

PubMed: 22840773
DOI: 10.1016/j.chembiol.2012.05.016

Experimental method

X-RAY DIFFRACTION (2.332 Å)