A versatile cis-prenyltransferase from Methanosarcina mazei catalyzes both C- and O-prenylations.

Polyprenyl groups, products of isoprenoid metabolism, are utilized in peptidoglycan biosynthesis, protein N-glycosylation, and other processes. These groups are formed by cis-prenyltransferases, which use allylic prenyl pyrophosphates as prenyl-donors to catalyze the C-prenylation of the general acceptor substrate, isopentenyl pyrophosphate. Repetition of this reaction forms (Z,E-mixed)-polyprenyl pyrophosphates, which are converted later into glycosyl carrier lipids, such as undecaprenyl phosphate and dolichyl phosphate. MM_0014 from the methanogenic archaeon Methanosarcina mazei is known as a versatile cis-prenyltransferase that accepts both isopentenyl pyrophosphate and dimethylallyl pyrophosphate as acceptor substrates. To learn more about this enzyme's catalytic activity, we determined the X-ray crystal structures of MM_0014 in the presence or absence of these substrates. Surprisingly, one structure revealed a complex with O-prenylglycerol, suggesting that the enzyme catalyzed the prenylation of glycerol contained in the crystallization buffer. Further analyses confirmed that the enzyme could catalyze the O-prenylation of small alcohols, such as 2-propanol, expanding our understanding of the catalytic ability of cis-prenyltransferases.

A heteromeric cis-prenyltransferase is responsible for the biosynthesis of glycosyl carrier lipids in Methanosarcina mazei.

Cis-prenyltransferases are enzymes responsible for the biosynthesis of glycosyl carrier lipids, natural rubber, and some secondary metabolites. Certain organisms, including some archaeal species, possess multiple genes encoding cis-prenyltransferase homologs, and the physiological roles of these seemingly-redundant genes are often obscure. Cis-prenyltransferases usually form homomeric complexes, but recent reports have demonstrated that certain eukaryotic enzymes are heteromeric protein complexes consisting of two homologous subunits. In this study, three cis-prenyltransferase homolog proteins, MM_0014, MM_0618, and MM_1083, from the methanogenic archaeon Methanosarcina mazei are overexpressed in Escherichia coli and partially purified for functional characterization. Coexistence of MM_0618 and MM_1083 exhibits prenyltransferase activity, while each of them alone has almost no activity. The chain-lengths of the products of this heteromeric enzyme are in good agreement with those of glycosyl carrier lipids extracted from M. mazei, which are likely di- and tetra-hydrogenated decaprenyl phosphates, suggesting that the MM_0618/MM_1083 heteromer is involved in glycosyl carrier lipid biosynthesis. MM_0014 acts as a typical homomeric cis-prenyltransferase and produces shorter products.

A cis-prenyltransferase from Methanosarcina acetivorans catalyzes both head-to-tail and nonhead-to-tail prenyl condensation.

Cis-prenyltransferase usually consecutively catalyzes the head-to-tail condensation reactions of isopentenyl diphosphate to allylic prenyl diphosphate in the production of (E,Z-mixed) polyprenyl diphosphate, which is the precursor of glycosyl carrier lipids. Some recently discovered homologs of the enzyme, however, catalyze the nonhead-to-tail condensation reactions between allylic prenyl diphosphates. In this study, we characterize a cis-prenyltransferase homolog from a methanogenic archaeon, Methanosarcina acetivorans, to obtain information on the biosynthesis of the glycosyl carrier lipids within it. This enzyme catalyzes both head-to-tail and nonhead-to-tail condensation reactions. The kinetic analysis shows that the main reaction of the enzyme is consecutive head-to-tail prenyl condensation reactions yielding polyprenyl diphosphates, while the chain lengths of the major products seem shorter than expected for the precursor of glycosyl carrier lipids. On the other hand, a subsidiary reaction of the enzyme, i.e., nonhead-to-tail condensation between dimethylallyl diphosphate and farnesyl diphosphate, gives a novel diterpenoid compound, geranyllavandulyl diphosphate.