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

Abstract

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.

Lipid sugar carriers at the extremes: The phosphodolichols Archaea use in N-glycosylation

“…In S. acidocaldarius , apparently only trans -geranylgeranyl pyrophosphate serves this role [92]. The recent report of one of the three cis -prenyltransferases in Methanosarcina acetivorans being able to catalyze both head-to-tail and non-head-to-tail condensation reactions [105] could reflect yet another unique aspect of archaeal phosphodolichol biosynthesis. In the hyperthermophile Aeropyrum pernix (optimal growth at 90–95°C [106]), the cis -prenyltransferase is unusual in that the substrate is the C 25 trans -geranylfarnesyl pyrophosphate [107].…”

“…In plants, where cis -prenyltransferases exist as enzyme families, it was shown that select family members that contribute to dolichol biogenesis also interact with a NogoB receptor homologue [103,110]. Although phylogenetic analysis has raised the possibility of such a protein in Archaea [105], evidence for the involvement of a NogoB receptor homologue in archaeal cis -prenyltransferase activity has yet to be presented.…”

Genome-wide gene expression and RNA half-life measurements allow predictions of regulation and metabolic behavior in Methanosarcina acetivorans

“…Fifth, metabolic pathways for incorporating the methylated sulfur compound, methylmercaptopropionate, have been added/updated based on recent molecular biology studies [57]. Finally, several genes and a recently characterized reaction have been added to lipid biosythesis [58–60]. …”

Purification of the mammalian NgBR/hCITcis-prenyltransferase complex: Identification of a conserved carboxyterminal RxG motif crucial for enzymatic activity

“…At first glance, this may appear to be in conflict with previous reports showing that eukaryotic hCIT orthologs heterologously expressed in S. cerevisiae were active without co-expression of NgBR/Nus1 orthologs. However, this can be explained due to the presence of the endogenous Nus1 in S. cerevisiae (7,10,(33)(34)(35)(37)(38)(39)(40). Furthermore, a number of hCIT orthologs in plants and protists that were studied in yeast are single-subunit enzymes that have been acquired through horizontal gene transfer, and are more closely related to EcUPPS than to hCIT (1,41).…”