Alternative oxidase: what information can protein sequence comparisons give us?
The finding that alternative oxidase (AOX) is present in most kingdoms of life has resulted in a large number of AOX sequences that are available for analyses. Multiple sequence alignments of AOX proteins from evolutionarily divergent organisms represent a valuable tool and can be used to identify amino acids and domains that may play a role in catalysis, membrane association and post-translational regulation, especially when these data are coupled with the structural model for the enzyme. I validate the use of this approach by demonstrating that it detects the conserved glutamate and histidine residues in AOX that initially led to its identification as a di-iron carboxylate protein and the generation of a structural model for the protein. A comparative analysis using a larger dataset identified 35 additional amino acids that are conserved in all AOXs examined, 30 of which have not been investigated to date. I hypothesize that these residues will be involved in the quinol terminal oxidase activity or membrane association of AOX. Major differences in AOX protein sequences between kingdoms are revealed, and it is hypothesized that two angiosperm-specific domains may be responsible for the non-covalent dimerization of AOX, whereas two indels in the aplastidic AOXs may play a role in their post-translational regulation. A scheme for predicting whether a particular AOX protein will be recognized by the alternative oxidase monoclonal antibody generated against the AOX of Sauromatum guttatum (Voodoo lily) is presented. The number of functional sites in AOX is greater than expected, and determining the structure of AOX will prove extremely valuable to future research.
McDonald
Allison E
AE
Department of Biology, The University of Western Ontario, 1151 Richmond St. N., London, Ontario N6A5B7, Canada. amcdon27@uwo.ca
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000276:immunology; D002462:Cell Membrane / Q000201:enzymology; D017124:Conserved Sequence; D024101:Mitochondrial Proteins; D008967:Molecular Biology; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry* / Q000276:immunology; D010940:Plant Proteins; D011499:Protein Processing, Post-Translational; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D000099188:Alternative Oxidase
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D000099188:Alternative Oxidase