Molten globule intermediates of human serum albumin in low concentration of urea.
Interaction of non-electrolytes such as urea with proteins especially at lower concentrations is opening-up newer concepts in the understanding of protein stability and folding in proteomics. In this study, the secondary and tertiary structural characteristics and thermal stability of human serum albumin at lower concentrations of urea have been monitored. The protein attains a molten globule like structure at concentration urea below 2 M. This structural state also shows an increase in the alpha-helical content as compared to the native state. At concentrations of urea above 2 M, human serum albumin starts unfolding, resulting in a three-state transition with two mid points of transitions at around 4 M and 7 M urea concentrations. The characteristics of the partially folded intermediates are discussed with respect to the three component system analyses. Preferential hydration dominates over preferential interaction at lower concentration of urea (up to 2.5 M) and at higher concentration, the preferential interaction overtakes preferential hydration in a competitive manner. Formation of structural intermediates at lower concentration of urea is hypothesized as a general phenomenon in proteins and fits in with the observation with preferential interaction parameters by Timasheff and co-workers in the case of lysozyme and ribonuclease at different pH values.
Department of Protein Chemistry and Technology, Central Food Technological Research Institute, Mysore 570 013, India.
B K
BK
Muralidhara
V
V
Prakash
D002152:Calorimetry, Differential Scanning / Q000379:methods; D002942:Circular Dichroism; D004220:Disulfides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000737:chemistry; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013696:Temperature; D013816:Thermodynamics; D014508:Urea / Q000737:chemistry*
D004220:Disulfides; D011506:Proteins; D012709:Serum Albumin; D014508:Urea; D012260:Ribonucleases; D009113:Muramidase