Structure and Mechanism of a Eukaryotic FMN Adenylyltransferase

Carlos Huerta; Dominika Borek; Mischa Machius; Nick V. Grishin; Hong Zhang

doi:10.1016/j.jmb.2009.04.022

Structure and Mechanism of a Eukaryotic FMN Adenylyltransferase

Carlos Huerta, Dominika Borek, Mischa Machius, Nick V. Grishin, Hong Zhang

Research output: Contribution to journal › Article › peer-review

45 Scopus citations

Abstract

Flavin mononucleotide adenylyltransferase (FMNAT) catalyzes the formation of the essential flavocoenzyme flavin adenine dinucleotide (FAD) and plays an important role in flavocoenzyme homeostasis regulation. By sequence comparison, bacterial and eukaryotic FMNAT enzymes belong to two different protein superfamilies and apparently utilize different sets of active-site residues to accomplish the same chemistry. Here we report the first structural characterization of a eukaryotic FMNAT from the pathogenic yeast Candida glabrata. Four crystal structures of C. glabrata FMNAT in different complexed forms were determined at 1.20-1.95 Å resolutions, capturing the enzyme active-site states prior to and after catalysis. These structures reveal a novel flavin-binding mode and a unique enzyme-bound FAD conformation. Comparison of the bacterial and eukaryotic FMNATs provides a structural basis for understanding the convergent evolution of the same FMNAT activity from different protein ancestors. Structure-based investigation of the kinetic properties of FMNAT should offer insights into the regulatory mechanisms of FAD homeostasis by FMNAT in eukaryotic organisms.

Original language	English (US)
Pages (from-to)	388-400
Number of pages	13
Journal	Journal of Molecular Biology
Volume	389
Issue number	2
DOIs	https://doi.org/10.1016/j.jmb.2009.04.022
State	Published - Jun 5 2009

Keywords

FAD biosynthesis
Rossmann-like fold
adenylyltransferase
convergent evolution
flavocoenzymes

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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10.1016/j.jmb.2009.04.022

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title = "Structure and Mechanism of a Eukaryotic FMN Adenylyltransferase",

abstract = "Flavin mononucleotide adenylyltransferase (FMNAT) catalyzes the formation of the essential flavocoenzyme flavin adenine dinucleotide (FAD) and plays an important role in flavocoenzyme homeostasis regulation. By sequence comparison, bacterial and eukaryotic FMNAT enzymes belong to two different protein superfamilies and apparently utilize different sets of active-site residues to accomplish the same chemistry. Here we report the first structural characterization of a eukaryotic FMNAT from the pathogenic yeast Candida glabrata. Four crystal structures of C. glabrata FMNAT in different complexed forms were determined at 1.20-1.95 {\AA} resolutions, capturing the enzyme active-site states prior to and after catalysis. These structures reveal a novel flavin-binding mode and a unique enzyme-bound FAD conformation. Comparison of the bacterial and eukaryotic FMNATs provides a structural basis for understanding the convergent evolution of the same FMNAT activity from different protein ancestors. Structure-based investigation of the kinetic properties of FMNAT should offer insights into the regulatory mechanisms of FAD homeostasis by FMNAT in eukaryotic organisms.",

keywords = "FAD biosynthesis, Rossmann-like fold, adenylyltransferase, convergent evolution, flavocoenzymes",

author = "Carlos Huerta and Dominika Borek and Mischa Machius and Grishin, {Nick V.} and Hong Zhang",

note = "Funding Information: We thank Zbyszek Otwinowski for help with synchrotron data collection and initial SAD phasing. This work was supported by a grant from the Welch Foundation (I-5105). C.H. was supported by National Institutes of Health training grant T32 GM008297. Results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at the Advanced Photon Source. Argonne National Laboratory is operated by UChicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research, under contract no. DE-AC02-06CH11357. ",

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AU - Zhang, Hong

N1 - Funding Information: We thank Zbyszek Otwinowski for help with synchrotron data collection and initial SAD phasing. This work was supported by a grant from the Welch Foundation (I-5105). C.H. was supported by National Institutes of Health training grant T32 GM008297. Results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at the Advanced Photon Source. Argonne National Laboratory is operated by UChicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research, under contract no. DE-AC02-06CH11357.

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N2 - Flavin mononucleotide adenylyltransferase (FMNAT) catalyzes the formation of the essential flavocoenzyme flavin adenine dinucleotide (FAD) and plays an important role in flavocoenzyme homeostasis regulation. By sequence comparison, bacterial and eukaryotic FMNAT enzymes belong to two different protein superfamilies and apparently utilize different sets of active-site residues to accomplish the same chemistry. Here we report the first structural characterization of a eukaryotic FMNAT from the pathogenic yeast Candida glabrata. Four crystal structures of C. glabrata FMNAT in different complexed forms were determined at 1.20-1.95 Å resolutions, capturing the enzyme active-site states prior to and after catalysis. These structures reveal a novel flavin-binding mode and a unique enzyme-bound FAD conformation. Comparison of the bacterial and eukaryotic FMNATs provides a structural basis for understanding the convergent evolution of the same FMNAT activity from different protein ancestors. Structure-based investigation of the kinetic properties of FMNAT should offer insights into the regulatory mechanisms of FAD homeostasis by FMNAT in eukaryotic organisms.

AB - Flavin mononucleotide adenylyltransferase (FMNAT) catalyzes the formation of the essential flavocoenzyme flavin adenine dinucleotide (FAD) and plays an important role in flavocoenzyme homeostasis regulation. By sequence comparison, bacterial and eukaryotic FMNAT enzymes belong to two different protein superfamilies and apparently utilize different sets of active-site residues to accomplish the same chemistry. Here we report the first structural characterization of a eukaryotic FMNAT from the pathogenic yeast Candida glabrata. Four crystal structures of C. glabrata FMNAT in different complexed forms were determined at 1.20-1.95 Å resolutions, capturing the enzyme active-site states prior to and after catalysis. These structures reveal a novel flavin-binding mode and a unique enzyme-bound FAD conformation. Comparison of the bacterial and eukaryotic FMNATs provides a structural basis for understanding the convergent evolution of the same FMNAT activity from different protein ancestors. Structure-based investigation of the kinetic properties of FMNAT should offer insights into the regulatory mechanisms of FAD homeostasis by FMNAT in eukaryotic organisms.

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KW - convergent evolution

KW - flavocoenzymes

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Structure and Mechanism of a Eukaryotic FMN Adenylyltransferase

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