Thermal stability of human albumin measured by differential scanning calorimetry. I. Effects of caprylate and N-acetyltryptophanate.
The thermal stability of 5% previously unheated, undefatted human albumin monomer in 145 mM Na+, pH 7.0 was investigated by differential scanning calorimetry (DSC) as a function of added caprylate and/or N-acetyl-DL-tryptophanate. Caprylate was substantially more effective than N-acetyl-DL-tryptophanate in protecting the protein against thermal denaturation at a given level or at a saturating level of stabilizer. The tracing of the differential heat capacity versus temperature (thermogram) for this undefatted monomer that contained 1.5 mol endogenous, long-chain fatty acid (LCFA)/mol monomer exhibits two denaturation peaks (endotherms) in the absence of stabilizer. The endotherm with the lower denaturation temperature (Td) comprises 70% of the total heat of denaturation and also corresponds to irreversible denaturation and precipitation of 70% of the albumin. This endotherm is associated with more thermally labile protein species containing low levels of LCFA. The endotherm with the higher Td is associated with more stable protein species containing high levels of LCFA. Thus, the two endotherms are not related to the proposed domain structure of the protein but result from an uneven LCFA distribution that is due to preexisting heterogeneity in the albumin and/or heterogeneity that arises during the DSC experiment. Binding data do not support a preexisting uneven distribution of sufficient magnitude to explain the experimental results. A complementary explanation is that an uneven fatty acid distribution arises during the DSC experiment by migration of LCFA from the more labile species to the more stable as the former unfold; such migration would cause further stabilization of the latter.
D002152:Calorimetry, Differential Scanning; D002210:Caprylates; D005227:Fatty Acids; D006358:Hot Temperature; D006801:Humans; D011487:Protein Conformation; D011489:Protein Denaturation; D012709:Serum Albumin; D014364:Tryptophan / Q000031:analogs & derivatives
D002210:Caprylates; D005227:Fatty Acids; D012709:Serum Albumin; C006392:N-acetyltryptophan; D014364:Tryptophan