Artemin is an RNA-binding protein with high thermal stability and potential RNA chaperone activity.
Encysted embryos of the crustacean, Artemia franciscana, are among the most stress-resistant of all multicellular eukaryotes, due in part to massive amounts of p26, a small heat shock protein, that acts as a molecular chaperone. These embryos contain equally large amounts of another protein called artemin, of previously unknown function, that we report on here. Its thermal stability allows large-scale purification in about a day, using ammonium sulfate fractionation and incubation at 70 degrees C for 7 min, followed by gel filtration. The latter yields an artemin-RNA complex from which the pure protein, apo-artemin, was obtained by anion-exchange chromatography. We evaluated the possibility that artemin acts as a molecular chaperone for proteins, but obtained no evidence for that in vitro. The association of RNA with apo-artemin occurs at high temperatures and, although it is not yet clear whether artemin has a specific role as an RNA chaperone, it does bind non-polyadenylated RNAs which are then translated in vitro. Artemin-RNA is thermostable, some molecules resisting destruction after 30 min at 90 degrees C. The first order rate constant for denaturation and aggregation of artemin-RNA at 85 degrees C is 8.5 x 10(-3)min(-1), which compares well with other thermostable proteins of similar size ( approximately 500 kDa) such as the ferritins with which artemin has amino acid sequence similarity. The amount of artemin extracted from embryos that had been stored dry, under laboratory conditions, since 1951 is comparable to the amount in contemporary embryos, indicating its stability in situ, and supporting the in vitro heating studies.
Warner
A H
AH
Department of Biological Sciences, University of Windsor, Windsor, Ont., Canada N9B 3P4.
Brunet
R T
RT
MacRae
T H
TH
Clegg
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins; D002352:Carrier Proteins; D003445:Crustacea / Q000196:embryology* / Q000378:metabolism*; D004355:Drug Stability; D033862:Iron-Binding Proteins; D007700:Kinetics; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D011485:Protein Binding; D011489:Protein Denaturation; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry*; D013696:Temperature
D060829:Arthropod Proteins; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; C490888:artemin protein, brine shrimp; D012313:RNA