Masked cerulenin enables a dual-site selective protein crosslink

Area of Science:

• Natural Product Chemistry
• Chemical Biology
• Enzymology

Background:

• Protein-reactive natural products, like cerulenin, are valuable therapeutic candidates due to their selective reactivity with protein active sites.
• Fatty-acid synthases (FASs) and polyketide synthases (PKSs) are crucial enzymes in biosynthesis, but studying their activity and structure remains challenging.

Purpose of the Study:

• To develop novel chemical probes based on cerulenin for studying FAS and PKS enzymes.
• To adapt cerulenin's reactivity for fluorescent labeling and crosslinking of these enzyme classes.
• To create new masked crosslinking probes for advanced activity and structure investigations of biosynthetic pathways.

Main Methods:

• Synthetic modulation of cerulenin's functional moieties to alter its reactivity.
• Mechanism-based investigation of cerulenin's unique chemical properties.
• Development of fluorescent labeling and crosslinking strategies using modified cerulenin.
• Design and synthesis of silylcyanohydrin and silylhemiaminal masked crosslinking probes.

Main Results:

• Successful synthetic modification of cerulenin yielded probes with tailored reactivity.
• Demonstrated utility of modified cerulenin for fluorescent labeling and crosslinking of fatty-acid and iterative type-I polyketide synthases.
• Introduction of two novel classes of masked crosslinking probes, expanding the toolkit for biochemical studies.

Conclusions:

• Modified cerulenin derivatives serve as effective tools for investigating FAS and PKS enzymes.
• The developed probes facilitate fluorescent labeling, crosslinking, and structural studies of these critical biosynthetic pathways.
• New masked crosslinking probes offer enhanced capabilities for exploring enzyme activity and mechanisms.