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NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP

Nature volume 401, pages 818–822 (1999)Cite this article

Abstract

The inhibitor-of-apoptosis (IAP) family of proteins, originally identified in baculoviruses1, regulate programmed cell death in a variety of organisms2,3,4,5,6. IAPs inhibit specific enzymes (caspases) in the death cascade7,8,9,10,11 and contain one to three modules of a common 70-amino-acid motif called the BIR domain12. Here we describe the nuclear magnetic resonance structure of a region encompassing the second BIR domain (BIR2) of a human IAP family member, XIAP (also called hILP or MIHA). The structure of the BIR domain consists of a three-stranded antiparallel β-sheet and four α-helices and resembles a classical zinc finger13. Unexpectedly, conserved amino acids within the linker region between the BIR1 and BIR2 domains were found to be critical for inhibiting caspase-3. The absence or presence of these residues may explain the differences in caspase inhibition observed for different truncated and full-length IAPs10,11. Our data further indicate that these residues may bind to the active site and that the BIR domain may interact with an adjacent site on the enzyme.

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Figure 1: Structure of the BIR2 domain of XIAP.
Figure 2: Surface of the BIR2 domain of XIAP.
Figure 3: Binding of (C202A/C213G)XIAP (residues 124–240) to His-tagged caspase-3 in the presence and absence of the caspase-3 inhibitor DEVD aldehyde.

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Acknowledgements

We thank R. Mendoza for technical assistance; A. Pardi for the starter cultures of Pseudomonas aeruginosa and Pf1 phage; M. Clore for a modified version of the X-PLOR program; F. Delaglio and A. Bax for the TALOS program; and J. Wu from IDUN Pharmaceuticals for the caspase-3 clone.

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Author notes

  1. Chaohong Sun and Mengli Cai: These authors contributed equally to this work

  2. Stephen W. Fesik: Correspondence and requests for materials should be addressed to S.W.F. Coordinates for the NMR structure of (C202A/C213G)XIAP (residues 124–240) have been deposited in the Brookhaven Protein Data Bank under the accession code 1C9Q.

Authors and Affiliations

  1. Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, 60064, Illinois, USA

    Chaohong Sun, Mengli Cai, Angelo H. Gunasekera, Robert P. Meadows, Hong Wang, Jun Chen, Haichao Zhang, Wei Wu, Nan Xu, Shi-Chung Ng & Stephen W. Fesik

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Correspondence to Stephen W. Fesik.

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Sun, C., Cai, M., Gunasekera, A. et al. NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP. Nature 401, 818–822 (1999). https://doi.org/10.1038/44617

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