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Sebastiaan Werten

Institute for General, Inorganic and Theoretical Chemistry, University of Innsbruck, Innrain 80-82, Innsbruck, 6020, Austria.

4 papers in the library · 49 citations · publishing 2022-2024

Papers

Methyl transfer in psilocybin biosynthesis

Nature Communications March 28, 2024 Jesse Hudspeth, Kai Rogge, Sebastian Dörner et al. 24 citations

Psilocybin, the natural hallucinogen in magic mushrooms, is produced in a final biosynthetic step where the enzyme PsiM adds two methyl groups to norbaeocystin. Atomic-resolution crystal structures (0.9 Å) of PsiM at different reaction stages reveal its detailed methylation mechanism. Structural and phylogenetic evidence indicates PsiM evolved from METTL16-family RNA methyltransferases, and its bound substrates mimic RNA. Limitations inherited from its ancestral scaffold prevent efficient psilocybin assembly and block trimethylation to aeruginascin. These insights will aid bioengineering efforts to create psilocybin variants with improved therapeutic properties.

Characterization of the Gateway Decarboxylase for Psilocybin Biosynthesis

ChemBioChem November 3, 2022 Tim Schäfer, Kristina Kramer, Sebastiaan Werten et al. 20 citations

The enzyme PsiD from the mushroom Psilocybe cubensis initiates the production of psilocybin, the psychedelic compound in magic mushrooms that is being investigated as a treatment for major depressive disorder. Unlike most similar enzymes, PsiD does not require pyridoxal phosphate (PLP) and instead resembles type II phosphatidylserine decarboxylases. Through biochemical experiments and computer modeling, researchers identified and confirmed a non-standard serine protease triad that enables the enzyme to cleave itself into its active form. This finding clarifies the molecular mechanism behind the first step of psilocybin biosynthesis.

Substrate recognition by the 4‐hydroxytryptamine kinase PsiK in psilocybin biosynthesis

FEBS Letters October 24, 2024 Kai Rogge, Tobias Wagner, Dirk Hoffmeister et al. 4 citations

Psilocybin, the hallucinogen from magic mushrooms, is being developed as a treatment for depression and other mental health conditions. Its biosynthesis from the amino acid L-tryptophan requires four sequential steps, the third of which is ATP-dependent phosphorylation of the intermediate 4-hydroxytryptamine, catalyzed by the enzyme PsiK. A crystallographic analysis and structure-based mutagenesis study of PsiK reveals how it recognizes its substrate. These findings will aid future bioengineering to create psilocybin variants with improved therapeutic properties.

The Second Methylation in Psilocybin Biosynthesis Is Enabled by a Hydrogen Bonding Network Extending into the Secondary Sphere Surrounding the Methyltransferase Active Site

ChemBioChem October 16, 2024 Jesse Hudspeth, Kai Rogge, Tobias Wagner et al. 1 citation

The enzyme PsiM from the mushroom Psilocybe cubensis catalyzes the final step in psilocybin biosynthesis, adding two methyl groups to the substrate norbaeocystin. A single amino acid change, M247N, allowed this enzyme to evolve from ancestral monomethylating RNA methyltransferases into a dimethylating enzyme. Mutating this asparagine back to methionine (N247M) or alanine (N247A) eliminated the ability to perform the second methylation. High-resolution crystal structures and kinetic measurements show that Asn247 provides necessary space in the active site for multiple methylations and stabilizes nearby secondary structures through hydrogen bonds, enabling efficient substrate binding and catalysis.