Abstract
Pressure unfolding–refolding and the subsequent aggregation of human serum albumin (HSA) was investigated by high-pressure Fourier transform infrared measurements. HSA is completely unfolded at 1 GPa pressure, but the unfolding is not cooperative. Hydrogen–deuterium exchange experiments suggest that a molten globule-like conformation is adopted above 0.4 GPa. An intermediate was formed after decompression, which differs from the native state only slightly in terms of the secondary structure, but this intermediate is more stable against the temperature-induced gel formation than the pressure-untreated native protein. This observation can be explained by assuming that the pressure unfolded–refolded protein is in a misfolded state, which is more stable than the native one.
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References
Booth DR, Sunde M, Bellotti V, Robinson CV, Hutchinson WL, Fraser PE, Hawkins FN, Dobson CM, Radford SE, Blake DDF, Pepys MB (1997) Instability, unfolding and aggregation of human lysozyme variants underlying amyoloid fibrillogenesis. Nature 385:787–793
Cordeiro Y, Kraineva J, Gomes MPB, Lopes MH, Martins VR, Lima LMTR, Foguel D, Winter R, Silva JL (2005) The amino-terminal PrP domain is crucial to modulate prion misfolding and aggregation. Biophys J 89:2667–2676
Cordeiro Y, Kraineva J, Suarez MC, Tempesta AG, Kelly JW, Silva JL, Winter R, Foguel D (2006) Fourier transform infrared spectroscopy provides a fingerprint for the tetramer and for the aggregates of transthyretin. Biophys J 91:957–967
Daggett V, Fersht AR (2003) Is there a unifying mechanism for protein folding? Trends Biochem Sci 28:18–25
Dobson CM (2003) Protein folding and misfolding. Nature 426:884–890
Dobson CM (2004) Principles of protein folding, misfolding and aggregation. Semin Cell Dev Biol 15:3–16
Dumoulin D, Last AM, Desmyter A, Decanniere K, Canet D, Larsson G, Spencer A, Archer DB, Sasse J, Muyldermans S, Wyns L, Redfield C, Matagne A, Robinson CV, Dobson CM (2003) A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme. Nature 424:783–787
Englander SW, Kallenbach NR (1984) Hydrogen exchange and structural dynamics of proteins and nucleic acids. Q Rev Biophys 16:521–655
Foguel D, Suarez MC, Ferrão-Gonzales AD, Porto TCR, Palmieri L, Einsiedler CM, Andrade LR, Lashuel HA, Lansbury PT, Kelly JW, Silva JL (2003) Dissociation of amyloid fibrils of alpha-synuclein and transthyretin by pressure reveals their reversible nature and the formation of water-excluded cavities. Proc Natl Acad Sci USA 100:9831–9836
Goosens K, Smeller L, Frank J, Heremans K (1996) Conformation of bovine pancretic trypsin inhibitor studied by fourier transform infrared spectroscopy. Eur J Biochem 236:254–262
Gruebele M (1999) The fast protein folding problem. Annu Rev Phys Chem 50:485–516
Grudzielanek S, Smirnovas V, Winter R (2006) Title: Solvation-assisted pressure tuning of insulin fibrillation: From novel aggregation pathways to biotechnological applications. J Mol Biol 356:497–509
Harris PI, Chapman D (1995) The conformational analysis of peptides using Fourier transform IR spectroscopy. Biopolymers 34:251–263
He XM, Carter DC (1992) Atomic structure and chemistry of human serum albumin. Nature 358:209–215
Ismail AA, Mantsch HH, Wong PTT (1992) Aggregation of chymotrypsinogen portrait by FT-IR spectroscopy. Biochim Biophys Acta 1121:183–188
Kauppinen JK, Moffat DJ, Mantsch HH, Cameron DG (1981) Fourier-selfdeconvolution: A method for resolving intrinsically overlapped bands. Appl Spectrosc 35:271–276
Krishnakumar SS, Panda D (2002) Spatial relationship between the prodan site, Trp-214, and Cys-34 residues in human serum albumin and loss of structure through incremental unfolding. Biochemistry 41:7443–7452
Kumar M, Banerjee A, Rahaman O, Panda D (2005) Unfolding pathways of human serum albumin: evidence for sequential unfolding and folding of its three domains. Int J Biol Macromol 37:200–204
Meersman F, Smeller L, Heremans K (2002) A comparative FTIR study of cold-, pressure- and heat-induced unfolding and aggregation of myoglobin. Biophys J 82:2635–2644
Mozhaev VV, Heremans K, Frank J, Masson P, Balny C (1996) High-pressure effects on protein structure and function. Proteins Struct Funct Gen 24:81–91
Okuno A, Kato M, Taniguchi Y (2007) Pressure effects on the heat-induced aggregation of equine serum albumin by FT-IR spectroscopic study: secondary structure kinetic and thermodynamic properties. Biophys Biochim Acta 1774:652–660
Osváth S, Sabelko JJ, Gruebele M (2003) Tuning the heterogeneous early folding dynamics of phosphoglycerate kinase. J Mol Biol 333:187–199
Randolph TW, Seefeldt M, Carpenter JF (2002) High hydrostatic pressure as a tool to study protein aggreagion and amyloidosis. Biochim Biophys Acta 1595:224–234
Ruan K, Weber G (1989) Hysteresis and conformational drift of pressure-dissociated glyceraldehydephosphate dehydrogenase. Biochemistry 28:2144–2153
Smeller L (2002) Pressure–temperature phase diagram of biomolecules, Biochim Biophys Acta 1595:11–29
Smeller L, Goossens K, Heremans K (1995) How to avoid artifacts in Fourier self-deconvolution. Appl Spectrosc 49:1538–1542
Smeller L, Rubens P, Heremans K (1999) Pressure effect on the temperature induced unfolding and tendency to aggregate of myoglobin. Biochemistry 38:3816–3820
Smeller L, Meersman F, Fidy J, Heremans K (2003) High-pressure FTIR study of the stability of horseradish peroxidase. Effect of heme substitution, ligand binding, Ca++ removal and reduction of the disulfide bonds. Biochemistry 42:553–561
Smeller L, Meersman F, Heremans K (2006) Refolding studies using pressure. The energy landscape of lysozyme in the pressure–temperature plane. Biochim Biophys Acta Proteins Proteomics 1764:497–505
Sugio S, Kashima A, Mochizuki S, Noda M, Kobayashi K (1999) Crystal structure of human serum albumin at Angstrom resolution. Protein Eng 12:439–443
Susi H, Byler DM (1986) Resolution-enhanced Fourier-tranform infrared-spectroscopy of enzymes. Meth Enzymol 130:290–311
Tanaka N, Nishizawa H, Kunugi S (1997) Structure of pressure-induced denatured state of human serum albumin: a comparison with the intermediate in urea-induced denatruation. Biochim Biophys Acta 1338:13–20
Torrent J, Alvarez-Martinez MT, Harricane MC, Heitz F, Liautard JP, Balny C, Lange R (2004) High-pressure induces scrapie-like prion proteins misfolding and amyloid formation. Biochemistry 43:7162–7170
Wong PTT, Moffat DJ (1989) A new internal pressure calibrant for high-pressure infrared spectroscopy of aqueous systems. Appl Spectrosc 43:1279–1281
Acknowledgments
This work was supported by the Hungarian Science Fund OTKA49213 and the Hungarian-Flemish (MTA-FWO) exchange program. F.M. is a postdoctoral research fellow of the Research Foundation Flanders (FWO-Vlaanderen). The authors are grateful to the COST D30 Action (WG006-03).
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Regional Biophysics Conference of the National Biophysical Societies of Austria, Croatia, Hungary, Italy, Serbia and Slovenia.
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Smeller, L., Meersman, F. & Heremans, K. Stable misfolded states of human serum albumin revealed by high-pressure infrared spectroscopic studies. Eur Biophys J 37, 1127–1132 (2008). https://doi.org/10.1007/s00249-008-0277-0
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DOI: https://doi.org/10.1007/s00249-008-0277-0