The preaggregated state of an amyloidogenic protein: hydrostatic pressure converts native transthyretin into the amyloidogenic state.

Protein misfolding and aggregation cause several diseases, by mechanisms that are poorly understood. The formation of amyloid aggregates is the hallmark of most of these diseases. Here, the properties and formation of amyloidogenic intermediates of transthyretin (TTR) were investigated by the use of hydrostatic pressure and spectroscopic techniques. Native TTR tetramers (T(4)) were denatured by high pressure into a conformation that exposes tryptophan residues to the aqueous environment. This conformation was able to bind the hydrophobic probe bis-(8-anilinonaphthalene-1-sulfonate), indicating persistence of elements of secondary and tertiary structure. Lowering the temperature facilitated the pressure-induced denaturation of TTR, which suggests an important role of entropy in stabilizing the native protein. Gel filtration chromatography showed that after a cycle of compression-decompression at 1 degrees C, the main species present was a tetramer, with a small population of monomers. This tetramer, designated T(4)*, had a non-native conformation: it bound more bis-(8-anilinonaphthalene-1-sulfonate) than native T(4), was less stable under pressure, and on decompression formed aggregates under mild acidic conditions (pH 5-5.6). Our data show that hydrostatic pressure converts native tetramers of TTR into an altered state that shares properties with a previously described amyloidogenic intermediate, and it may be an intermediate that lies on the aggregation pathway. This "preaggregated" state, which we call T(4)*, provides insight into the question of how a correctly folded protein may degenerate into the aggregation pathway in amyloidogenic diseases.

[1]  F E Cohen,et al.  Pathologic conformations of prion proteins. , 1998, Annual review of biochemistry.

[2]  A. Jonas,et al.  High-pressure NMR spectroscopy of proteins and membranes. , 1994, Annual review of biophysics and biomolecular structure.

[3]  T. Baldwin,et al.  Kinetic partitioning during protein folding yields multiple native states , 1994, Nature Structural Biology.

[4]  J. Kelly,et al.  Guanidine hydrochloride-induced denaturation and refolding of transthyretin exhibits a marked hysteresis: equilibria with high kinetic barriers. , 1997, Biochemistry.

[5]  Maria João,et al.  Transthyretin mutations in health and disease , 1995, Human mutation.

[6]  J. Silva,et al.  Molten-globule conformation of Arc repressor monomers determined by high-pressure 1H NMR spectroscopy. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[7]  I. Dacklin,et al.  Characterization of two highly amyloidogenic mutants of transthyretin. , 1997, Biochemistry.

[8]  P. Lansbury,et al.  Models of amyloid seeding in Alzheimer's disease and scrapie: mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins. , 1997, Annual review of biochemistry.

[9]  D. Foguel,et al.  Pressure-induced Dissociation and Denaturation of Allophycocyanin at Subzero Temperatures (*) , 1995, Journal of Biological Chemistry.

[10]  J. Kelly,et al.  Comparison of lethal and nonlethal transthyretin variants and their relationship to amyloid disease. , 1995, Biochemistry.

[11]  G. Weber.  Phenomenological description of the association of protein subunits subjected to conformational drift. Effects of dilution and of hydrostatic pressure. , 1986, Biochemistry.

[12]  R. Canfield,et al.  Polymorphism of Rhesus monkey serum prealbumin. Purification and partial structure. , 1973, Journal of Biological Chemistry.

[13]  A. Fink,et al.  Protein conformational changes induced by 1,1'-bis(4-anilino-5-naphthalenesulfonic acid): preferential binding to the molten globule of DnaK. , 1994, Biochemistry.

[14]  J. Kelly,et al.  Transthyretin mutation Leu-55-Pro significantly alters tetramer stability and increases amyloidogenicity. , 1993, Biochemistry.

[15]  G. de Prat-Gay,et al.  Folding of a pressure-denatured model protein. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Kelly,et al.  Partial denaturation of transthyretin is sufficient for amyloid fibril formation in vitro. , 1992, Biochemistry.

[17]  J. Silva,et al.  Dissociation of a native dimer to a molten globule monomer. Effects of pressure and dilution on the association equilibrium of arc repressor. , 1992, Journal of molecular biology.

[18]  R. Wetzel.  Mutations and off-pathway aggregation of proteins. , 1994, Trends in biotechnology.

[19]  P. Prevelige,et al.  The use of hydrostatic pressure as a tool to study viruses and other macromolecular assemblages. , 1996, Current opinion in structural biology.

[20]  J. Jonas,et al.  Structure of pressure-assisted cold denatured lysozyme and comparison with lysozyme folding intermediates. , 1997, Biochemistry.

[21]  N. Bec,et al.  A stable partly denatured state of trypsin induced by high hydrostatic pressure. , 1997, Biochemical and biophysical research communications.

[22]  J. King,et al.  Thermolabile folding intermediates: inclusion body precursors and chaperonin substrates , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[23]  G. Hummer,et al.  The pressure dependence of hydrophobic interactions is consistent with the observed pressure denaturation of proteins. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  D. Foguel,et al.  Characterization of a Partially Folded Monomer of the DNA-binding Domain of Human Papillomavirus E2 Protein Obtained at High Pressure* , 1998, The Journal of Biological Chemistry.

[25]  Arthur L Horwich,et al.  Deadly Conformations—Protein Misfolding in Prion Disease , 1997, Cell.

[26]  J. Kelly,et al.  The acid-mediated denaturation pathway of transthyretin yields a conformational intermediate that can self-assemble into amyloid. , 1996, Biochemistry.

[27]  A. Jonas,et al.  NMR study of the cold, heat, and pressure unfolding of ribonuclease A. , 1995, Biochemistry.

[28]  R. Sauer,et al.  Formation of a denatured dimer limits the thermal stability of Arc repressor. , 1997, Journal of molecular biology.

[29]  J. Silva,et al.  Arc repressor will not denature under pressure in the absence of water. , 1994, Journal of molecular biology.

[30]  Jerson L. Silva,et al.  Pressure stability of proteins. , 1993, Annual review of physical chemistry.

[31]  J. Kelly,et al.  The alternative conformations of amyloidogenic proteins and their multi-step assembly pathways. , 1998, Current opinion in structural biology.

[32]  J. Carpenter,et al.  High pressure fosters protein refolding from aggregates at high concentrations. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[33]  S J Oatley,et al.  Structure of prealbumin: secondary, tertiary and quaternary interactions determined by Fourier refinement at 1.8 A. , 1977, Journal of molecular biology.

[34]  P. Lansbury,et al.  Amyloid diseases: abnormal protein aggregation in neurodegeneration. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[35]  R. Brito,et al.  The amyloidogenic potential of transthyretin variants correlates with their tendency to aggregate in solution , 1997, FEBS letters.

[36]  K. Heremans,et al.  Pressure effect on the temperature-induced unfolding and tendency to aggregate of myoglobin. , 1999, Biochemistry.

[37]  G. Glenner,et al.  THE RELATION OF THE PROPERTIES OF CONGO RED-STAINED AMYLOID FIBRILS TO THE β-CONFORMATION , 1972, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[38]  D. Foguel,et al.  Hydrostatic pressure rescues native protein from aggregates. , 1999, Biotechnology and bioengineering.

[39]  J. Liepnieks,et al.  Thyroxine interactions with transthyretin: a comparison of 10 different naturally occurring human transthyretin variants. , 1993, The Journal of clinical endocrinology and metabolism.

[40]  W. Kauzmann,et al.  Pressure dependence of weak acid ionization in aqueous buffers , 1973 .

[41]  R. Seckler,et al.  Formation of Fibrous Aggregates from a Non-native Intermediate: The Isolated P22 Tailspike β-Helix Domain* , 1999, The Journal of Biological Chemistry.

[42]  R. Seckler,et al.  In vitro folding pathway of phage P22 tailspike protein. , 1991, Biochemistry.

[43]  S. Cheng,et al.  Analysis of thyroid hormone binding to human serum prealbumin by 8-anilinonaphthalene-1-sulfonate fluorescence. , 1977, Biochemistry.

[44]  J. Kelly,et al.  Characterization of the transthyretin acid denaturation pathways by analytical ultracentrifugation: implications for wild-type, V30M, and L55P amyloid fibril formation. , 1998, Biochemistry.

[45]  C. Royer,et al.  Determination of the volume changes for pressure-induced transitions of apomyoglobin between the native, molten globule, and unfolded states. , 1998, Biophysical journal.

[46]  B Fane,et al.  Global suppression of protein folding defects and inclusion body formation. , 1991, Science.

[47]  K. Heremans,et al.  Protein structure and dynamics at high pressure. , 1998, Biochimica et biophysica acta.

[48]  Scott A. Peterson,et al.  Transthyretin quaternary and tertiary structural changes facilitate misassembly into amyloid. , 1997, Advances in protein chemistry.