Book of Abstracts: Albany 2009

category image Albany 2009
Conversation 16
June 16-20 2009
© Adenine Press (2008)

Structural Insights Into the Unusual Chemistry of Sir2 Enzymes

Sir2-like enzymes, also known as sirtuins, comprise a universally conserved family of NAD+-dependent protein deacetylases that play important roles in transcriptional silencing, fat mobilization, metabolic regulation, and lifespan extension. NAD+ is cleaved during the deacetylation reaction, yielding nicotinamide, deacetylated peptide and O-acetyl ADP-ribose products. In some cases, sirtuins catalyze mono-ADP ribosylation of their substrates rather than deacetylation. In an effort to determine the enzymatic mechanism of the NAD+-dependent deacetylation reaction, we have determined structures of sirtuins bound to a variety of substrates and products, as well as to a transition state analogue. In addition, we have trapped a key covalent reaction intermediate in complex with the enzyme and solved its structure. These structures provide insights into the structure-based mechanism of NAD+ cleavage and deacetylation. These mechanistic insights have been extended to explain the second reaction, ADP-ribosylation, that is catalyzed by some sirtuins. In solution studies of a trypanosomal sirtuin, we have identified the side chains that are ADP-ribosylated by TbSir2 and present a plausible mechanism for the dual activity based on our structural studies.

Cynthia Wolberger*
William Hawse
Kamau Fahie
Kevin Hoff

Dept of Biophysics
and Biophysical Chemistry
Howard Hughes Medical Inst
Johns Hopkins Univ.
School of Medicine
725 N. Wolfe St.
Baltimore, MD 21205

Phone: (410)955-0728
Fax: (410)614-8648