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Originally published In Press as doi:10.1074/jbc.M112045200 on January 18, 2002

J. Biol. Chem., Vol. 277, Issue 14, 12275-12279, April 5, 2002
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SMAC Negatively Regulates the Anti-apoptotic Activity of Melanoma Inhibitor of Apoptosis (ML-IAP)*

Domagoj VucicDagger , Kurt Deshayes§, Heidi Ackerly§, Maria Teresa Pisabarro§, Saloumeh Kadkhodayan, Wayne J. Fairbrother§, and Vishva M. DixitDagger ||

From the Departments of Dagger  Molecular Oncology, § Protein Engineering, and  Bioanalytical Research and Development, Genentech, Inc., South San Francisco, California 94080

Inhibitors of apoptosis (IAPs) physically interact with a variety of pro-apoptotic proteins and inhibit apoptosis induced by diverse stimuli. X-linked IAP (X-IAP) is a prototype IAP family member that inhibits several caspases, the effector proteases of apoptosis. The inhibitory activity of X-IAP is regulated by SMAC, a protein that is processed to its active form upon receipt of a death stimulus. Cleaved SMAC binds X-IAP and antagonizes its anti-apoptotic activity. Here we show that melanoma IAP (ML-IAP), a potent anti-cell death protein and caspase inhibitor, physically interacts with SMAC through its BIR (baculovirus IAP repeat) domain. In addition to binding full-length SMAC, ML-IAP BIR associates with SMAC peptides that are derived from the amino terminus of active, processed SMAC. This high affinity interaction is very specific and can be completely abolished by single amino acid mutations either in the amino terminus of active SMAC or in the BIR domain of ML-IAP. In cells expressing ML-IAP and X-IAP, SMAC coexpression or addition of SMAC peptides abrogates the ability of the IAPs to inhibit cell death. These results demonstrate the feasibility of using SMAC peptides as a way to sensitize IAP-expressing cells to pro-apoptotic stimuli such as chemotherapeutic agents.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed: Dept. of Molecular Oncology, Genentech, Inc., South San Francisco, CA 94080. Tel.: 650-225-1312; Fax: 650-225-6127; E-mail: dixit@gene.com.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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