Temperature behaviour of human serum albumin.
Structural alterations of albumin, their dependence on concentration and the role of free --SH groups at thermal denaturation, as well as the reversibility of thermally induced structural changes, were studied. Application of various physical methods provides information on a series of structural parameters in a major concentration range. Apart from changes of the helix content, heat treatment gives rise to beta structures which are amplified on cooling and which are correlated with the aggregation of albumin. With rising temperature and concentration the proportion of beta structures and aggregates increases. At degrees of denaturation of up to 20% complete renaturation is possibly in every case. The structure content is concentration-dependent even at room temperature. It may be that intermolecular interactions induce additional alpha-helix structures which are less stable, however, than the ones stabilized by intramolecular interactions. Unfolding of the pocket containing the free --SH group of cysteine-34 enables disulphide bridges to be formed leading to stable aggregates and irreversible structural alterations. Through binding of N-ethymaleimide to free --SH groups, which blocks the formation of disulphide bridges, it is possible to prevent aggregation and irreversible conformational changes. At temperatures below 65--70 degrees C, oligomers are formed mainly via intermolecular beta structures.
D002942:Circular Dichroism; D003545:Cysteine; D004220:Disulfides; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D011487:Protein Conformation; D011489:Protein Denaturation; D012709:Serum Albumin; D013055:Spectrophotometry, Infrared; D013696:Temperature; D014783:Viscosity
D004220:Disulfides; D012709:Serum Albumin; D003545:Cysteine