TY - JOUR

T1 - Biosynthesis and activity of prenylated FMN cofactors

AU - Wang, Po-Hsiang

AU - Khusnutdinova, Anna N.

AU - Luo, Fei

AU - Xiao, Johnny

AU - Nemr, Kayla

AU - Flick, Robert

AU - Brown, Greg

AU - Mahadevan, Radhakrishnan

AU - Edwards, Elizabeth

AU - Yakunin, Alexander

PY - 2018/5/17

Y1 - 2018/5/17

N2 - Prenylated FMN (prFMN) is a recently discovered cofactor required by the UbiD family of reversible decarboxylases involved in ubiquinone biosynthesis and in the biotransformation of aromatic compounds. This cofactor is synthesized by UbiX-like prenyltransferases catalyzing the transfer of the dimethylallyl moiety of dimethylallyl-monophosphate (DMAP) to FMN. The origin of DMAP for prFMN biosynthesis and the biochemical properties of free prFMN are unknown. We show that in Escherichia coli cells, DMAP can be produced by phosphorylating prenol using ThiM or dephosphorylating DMAPP using Nudix hydrolases. We produced 14 active prenyltransferases whose properties enabled the production of protein-free forms of prFMN. In vitro assays revealed that the UbiD-like ferulate decarboxylase (Fdc1) has high affinity to free prFMNiminium and C1′-ene-prFMNiminium and can be activated under both oxidized and reduced conditions. These insights into the biosynthesis and properties of prFMN will facilitate further elucidation of the biochemical diversity of reversible UbiD (de)carboxylases.

AB - Prenylated FMN (prFMN) is a recently discovered cofactor required by the UbiD family of reversible decarboxylases involved in ubiquinone biosynthesis and in the biotransformation of aromatic compounds. This cofactor is synthesized by UbiX-like prenyltransferases catalyzing the transfer of the dimethylallyl moiety of dimethylallyl-monophosphate (DMAP) to FMN. The origin of DMAP for prFMN biosynthesis and the biochemical properties of free prFMN are unknown. We show that in Escherichia coli cells, DMAP can be produced by phosphorylating prenol using ThiM or dephosphorylating DMAPP using Nudix hydrolases. We produced 14 active prenyltransferases whose properties enabled the production of protein-free forms of prFMN. In vitro assays revealed that the UbiD-like ferulate decarboxylase (Fdc1) has high affinity to free prFMNiminium and C1′-ene-prFMNiminium and can be activated under both oxidized and reduced conditions. These insights into the biosynthesis and properties of prFMN will facilitate further elucidation of the biochemical diversity of reversible UbiD (de)carboxylases.

U2 - 10.1016/j.chembiol.2018.02.007

DO - 10.1016/j.chembiol.2018.02.007

M3 - Article

VL - 25

SP - 560

EP - 570

JO - Cell Chemical Biology

JF - Cell Chemical Biology

SN - 2451-9448

IS - 5

ER -