title: Purification, structure and in vitro molecular-chaperone activity of Artemia p26, a small heat-shock/alpha-crystallin protein.
abstract: Encysted brine-shrimp gastrulae bring their metabolism to a reversible standstill during diapause and quiescence, demonstrating a remarkable resistance to unfavourable environmental conditions. For example, mortality of Artemia embryos under normal temperature and hydration is very low, even after two years of anoxia, and embryos commonly experience complete desiccation as part of their developmental program. Previous evidence from our laboratories indicated that p26, an abundant low-molecular-mass cyst-specific protein capable of translocation into the nucleus, may have a protective function in Artemia cysts. p26 was purified to apparent homogeneity and a continuous sequence of 141 of its amino acids was determined by peptide sequencing, revealing that it is a member of the small-heat-shock/alpha-crystallin family of proteins. As determined by molecular-sieve chromatography and sucrose-density-gradient centrifugation, native p26 is a multimer of about 27 monomers with a molecular mass of approximately 700 kDa. Inactivation of citrate synthase was less when the enzyme was heated in the presence rather than the absence of p26. Additionally, the renaturation of heat-inactivated citrate synthase was promoted by p26. These results indicated that p26 possesses molecular-chaperone activity, a property of other small heat-shock/alpha-crystallin proteins. Our findings demonstrate that p26 has the potential to protect the macromolecular components of Artemia embryos, either as they encyst or upon exposure to environmental extremes. Protection may depend upon the ability of p26 to function as a molecular chaperone.
authors.affiliation: Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: P
authors.initials: P
authors.lastname: Liang
authors.forename: R
authors.initials: R
authors.lastname: Amons
authors.forename: T H
authors.initials: TH
authors.lastname: Macrae
authors.forename: J S
authors.initials: JS
authors.lastname: Clegg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001156:Artemia / Q000737:chemistry* / Q000378:metabolism; D003459:Crystallins / Q000302:isolation & purification*; D003890:Desiccation; D006360:Heat-Shock Proteins / Q000302:isolation & purification*; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D003459:Crystallins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D018832:Molecular Chaperones