title: A highly conserved arginine is critical for the functional folding of inhibitor of apoptosis (IAP) BIR domains.
abstract: The inhibitor of apoptosis (IAP) proteins are found in all animals and regulate apoptosis (programmed cell death) by binding and inhibiting caspase proteases. This inhibition is overcome by several apoptosis stimulators, including Drosophila Hid and mammalian Smac/DIABLO, which bind to 65-residue baculovirus IAP repeat (BIR) domains found in one to three copies in all IAPs. Virtually all BIRs contain three Cys and a His that bind zinc, a Gly in a tight turn, and an Arg. The functional and structural role of the Arg was investigated in isolated BIR domains from the baculovirus Orgyia pseudotsugata Op-IAP and the Drosophila DIAP1 proteins. Mutation of the Arg to either Ala or Lys abolished Hid and Smac binding to BIRs, despite the Hid/Smac binding site being located on the opposite side of the BIR domain from the Arg. The mutant BIR domains also exhibited weakened zinc binding, increased sensitivity to limited proteolysis, and altered circular dichroism spectra indicative of perturbed domain folding. Examination of known BIR structures indicates that the Arg side chain makes simultaneous bridging hydrogen bonds and a cation-pi interaction for which the Arg guanidino group is uniquely well suited. These interactions are likely critical for stabilizing the tertiary fold of BIR domains in all IAPs, explaining the conservation of this residue.
authors.affiliation: Department of Molecular and Cell Biology, University of Texas at Dallas, P.O. Box 830688, Richardson, Texas 75083-0688, USA.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Luque
authors.forename: Katrina P
authors.initials: KP
authors.lastname: Grape
authors.forename: Matthew
authors.initials: M
authors.lastname: Junker
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001120:Arginine / Q000737:chemistry*; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D004330:Drosophila; D051034:Inhibitor of Apoptosis Proteins; D019476:Insect Proteins / Q000737:chemistry* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D017924:Nucleopolyhedroviruses; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D011506:Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D051034:Inhibitor of Apoptosis Proteins; D019476:Insect Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C086430:inhibitor of apoptosis, Nucleopolyhedrovirus; D001120:Arginine