The Structure of the Elusive Urease–Urea Complex Unveils the Mechanism of a Paradigmatic Nickel‐Dependent Enzyme

• Published in: Angewandte Chemie International Edition 2019-05, Vol.58 (22), p.7415-7419
• Main Authors: Mazzei, Luca, Cianci, Michele, Benini, Stefano, Ciurli, Stefano
• Format: Article
• Language: English
• Subjects: Catalysis; catalytic mechanism; Crystal structure; Enzymes; Fluorides; Nickel; Organic chemistry; Substrates; Urea; Ureas; Urease; x-ray diffraction
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201903565

Urease, the most efficient enzyme known, contains an essential dinuclear NiII cluster in the active site. It catalyzes the hydrolysis of urea, inducing a rapid pH increase that has negative effects on human health and agriculture. Thus, the control of urease activity is of utmost importance in medical, pharmaceutical, and agro‐environmental applications. All known urease inhibitors are either toxic or inefficient. The development of new and efficient chemicals able to inhibit urease relies on the knowledge of all steps of the catalytic mechanism. The short (microseconds) lifetime of the urease–urea complex has hampered the determination of its structure. The present study uses fluoride to substitute the hydroxide acting as the co‐substrate in the reaction, preventing the occurrence of the catalytic steps that follow substrate binding. The 1.42 Å crystal structure of the urease–urea complex, reported here, resolves the enduring debate on the mechanism of this metalloenzyme. Crystal clear: The crystal structure of the urease–urea complex, which reveals an unprecedented binding mode of urea to the di‐nickel cluster in the urease active site, provides definitive evidence for the initial step of the catalytic mechanism of urea hydrolysis. This finding could facilitate the development of improved urease inhibitors.