title: An exceptionally stable Group II chaperonin from the hyperthermophile Pyrococcus furiosus.
abstract: The hyperthermophilic archaeon Pyrococcus furiosus (Pf) grows optimally at 100 degrees C and encodes single genes for the Group II chaperonin (Cpn), Pf Cpn and alpha-crystallin homolog, the small Heat shock protein (sHsp). Recombinant Pf Cpn is exceptionally thermostable and remained active in high ionic strength, and up to 3M guanidine hydrochloride (Gdn-HCl). Pf Cpn bound specifically to denatured lysozyme and ATP addition resulted in protection of lysozyme from aggregation and inactivation at 100 degrees C. While complexed to heat inactivated lysozyme, Pf Cpn showed enhanced thermostability and ATPase activity, and increased the optimal temperature for ATPase activity from 90 to 100 degrees C. Protein substrate binding also stabilized the 16-mer oligomer of Pf Cpn in 3M Gdn-HCl and activated ATPase hydrolysis in 3-5M Gdn-HCl. In addition, Pf Cpn recognized and refolded the non-native lysozyme released from Pf sHsp, consistent with the inferred functions of these chaperones as the primary protein folding pathway during cellular heat shock.
authors.affiliation: Center of Marine Biotechnology, University of Maryland Institutes, 701 East Pratt Street, Baltimore, MD 21202, USA.
authors.forename: Haibin
authors.initials: H
authors.lastname: Luo
authors.forename: Pongpan
authors.initials: P
authors.lastname: Laksanalamai
authors.forename: Frank T
authors.initials: FT
authors.lastname: Robb
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D018833:Chaperonins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004355:Drug Stability; D019766:Genes, Archaeal; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D009113:Muramidase / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D020027:Pyrococcus furiosus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D019843:Archaeal Proteins; D017931:DNA Primers; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D009113:Muramidase; D000251:Adenosine Triphosphatases; D018833:Chaperonins