title: Truncation attenuates molecular chaperoning and apoptosis inhibition by p26, a small heat shock protein from Artemia franciscana.
abstract: The small heat shock proteins (sHSPs), which prevent irreversible protein denaturation and inhibit apoptosis, consist of an amino-terminus, the canonical α-crystallin domain, and a carboxy-terminal extension. It remains difficult, however, to define sHSP structure-function relationships and with this in mind p26, an sHSP from the crustacean Artemia franciscana, was truncated by deletion mutagenesis. Wild-type p26 cDNA and three truncated variants inserted into the eukaryotic expression vector pcDNA3.1/HisC were used to generate stably transfected 293H cells. p26 shielded transfected cells against death upon exposure to heat and oxidative stress. Truncation reduced chaperone activity, with cells synthesizing the p26 α-crystallin domain being the least resistant. Wild-type p26 inhibited apoptosis in transfected cells, with protection against oxidation-generated apoptosis being more effective than that against heat-induced apoptosis. Truncation reduced p26 apoptotic inhibitory activity, with the α-crystallin domain again being the least effective. The results show that a crustacean sHSP functions effectively in mammalian cells, demonstrating interchangeability of these proteins between distantly related organisms and indicating similarities in their mechanisms of action. Moreover, maximal activity was observed for full-length p26, indicating that structural elements required for chaperone activity and apoptosis inhibition reside throughout the protein.
authors.affiliation: Department of Biology, Dalhousie University, Halifax, NS, Canada.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wu
authors.forename: Thomas H
authors.initials: TH
authors.lastname: MacRae
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001156:Artemia / Q000235:genetics / Q000378:metabolism; D002461:Cell Line, Transformed; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D046188:Protein Modification, Translational; D017434:Protein Structure, Tertiary / Q000235:genetics; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D014162:Transfection; D038201:alpha-Crystallins / Q000235:genetics / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; D038201:alpha-Crystallins; C107632:p26 chaperone protein, Artemia franciscana