title: A kinetic assessment of the C. elegans amyloid disaggregation activity enables uncoupling of disassembly and proteolysis.
abstract: Protein aggregation is a common feature of late onset neurodegenerative disorders, including Alzheimer's disease. In Alzheimer's disease, misassembly of the Abeta peptide is genetically linked to proteotoxicity associated with disease etiology. A reduction in Abeta proteotoxicity is accomplished, in part, by the previously reported Abeta disaggregation and proteolysis activities-under partial control of heat shock factor 1, a transcription factor regulating proteostasis in the cytosol and negatively regulated by insulin growth factor signaling. Herein, we report an improved in vitro assay to quantify recombinant fibrillar Abeta disaggregation kinetics accomplished by the exogenous application of C.elegans extracts. With this assay we demonstrate that the Abeta disaggregation and proteolysis activities of C.elegans are separable. The disaggregation activity found in C.elegans preparations is more heat resistant than the proteolytic activity. Abeta disaggregation in the absence of proteolysis was found to be a reversible process. Future discovery of the molecular basis of the disaggregation and proteolysis activities offers the promise of delaying the age-onset proteotoxicity that leads to neurodegeneration in a spectrum of maladies.
authors.affiliation: Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Bieschke
authors.forename: Ehud
authors.initials: E
authors.lastname: Cohen
authors.forename: Amber
authors.initials: A
authors.lastname: Murray
authors.forename: Andrew
authors.initials: A
authors.lastname: Dillin
authors.forename: Jeffery W
authors.initials: JW
authors.lastname: Kelly
mesh_terms: D000544:Alzheimer Disease; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000737:chemistry* / Q000201:enzymology; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000378:metabolism*; D004195:Disease Models, Animal; D007700:Kinetics; D018625:Microscopy, Atomic Force; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D055503:Protein Multimerization; D013696:Temperature; D014020:Tissue Extracts / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D029742:Caenorhabditis elegans Proteins; D010446:Peptide Fragments; D014020:Tissue Extracts; C078592:amyloid beta-protein (1-40); D010447:Peptide Hydrolases
references.citation: J Struct Biol. 2005 Jan;149(1):30-7
references.pmid: 15629655
references.citation: Biophys J. 2006 Jul 1;91(1):122-32
references.pmid: 16603497
references.citation: Lancet. 2003 May 10;361(9369):1642-4
references.pmid: 12747895
references.citation: Science. 2002 Oct 25;298(5594):830-4
references.pmid: 12399591
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):4109-12
references.pmid: 9108113
references.citation: Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9368-72
references.pmid: 7568134
references.citation: Biochemistry. 2005 Apr 5;44(13):4977-83
references.pmid: 15794636
references.citation: Nature. 2003 Jan 9;421(6919):182-7
references.pmid: 12483226
references.citation: Mol Biol Cell. 2004 Feb;15(2):657-64
references.pmid: 14668486
references.citation: Science. 2006 Sep 15;313(5793):1604-10
references.pmid: 16902091
references.citation: Curr Opin Neurol. 1994 Aug;7(4):283-6
references.pmid: 7952234
references.citation: Adv Exp Med Biol. 2007;594:167-89
references.pmid: 17205684
references.citation: Methods Enzymol. 1999;309:274-84
references.pmid: 10507030
references.citation: Science. 2004 Jun 18;304(5678):1793-7
references.pmid: 15155912
references.citation: Curr Opin Struct Biol. 1998 Feb;8(1):101-6
references.pmid: 9519302
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3438-42
references.pmid: 18316725
references.citation: Nature. 1999 Jul 8;400(6740):173-7
references.pmid: 10408445
references.citation: Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10417-22
references.pmid: 12122205
references.citation: Brain Pathol. 1998 Jul;8(3):499-513
references.pmid: 9669700
references.citation: Cell. 2005 Feb 25;120(4):449-60
references.pmid: 15734678
references.citation: FEBS J. 2006 Apr;273(7):1331-49
references.pmid: 16689923
references.citation: Science. 2008 Feb 15;319(5865):916-9
references.pmid: 18276881
references.citation: J Mol Biol. 2007 Nov 9;373(5):1321-33
references.pmid: 17905305
references.citation: Cell. 2005 Feb 25;120(4):545-55
references.pmid: 15734686
references.citation: Nature. 2003 Dec 18;426(6968):884-90
references.pmid: 14685248
references.citation: Cell. 1998 Jul 10;94(1):73-82
references.pmid: 9674429
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 16;105(37):13987-92
references.pmid: 18765803
references.citation: Nat Rev Neurosci. 2008 Oct;9(10):759-67
references.pmid: 18769445
references.citation: Cell. 1997 Aug 8;90(3):549-58
references.pmid: 9267034
references.citation: Mol Cell. 2006 Aug 4;23(3):425-38
references.pmid: 16885031
references.citation: Cell. 2005 Apr 8;121(1):73-85
references.pmid: 15820680
references.citation: Neuron. 2003 Dec 18;40(6):1087-93
references.pmid: 14687544
references.citation: Nat Cell Biol. 2004 Nov;6(11):1054-61
references.pmid: 15516999
references.citation: Science. 2003 May 16;300(5622):1142-5
references.pmid: 12750521
references.citation: Nature. 1994 Dec 1;372(6505):475-8
references.pmid: 7984243