Stabilizing effects of caprylate and acetyltryptophanate on heat-induced aggregation of bovine serum albumin.
Acetyltryptophanate (AT) and caprylate (Cap) have been used to stabilize serum albumin against heat treatment. However, the mechanism of stabilization by these additives has never been fully elucidated. Here we used thermal melting to determine the effects of these additives on the melting temperature of bovine serum albumin (BSA) and heat stress at 60 degrees C to follow degradation of the protein in the presence of varying concentrations of AT or Cap. Native polyacrylamide gel electrophoresis was used to examine degradation products generated by heat treatment. Both additives increased the melting temperature of BSA, resulting in an increase by 12 degrees C at 5 mM AT and 3 degrees C at 1 mM Cap. They also conferred stability to BSA against heat stress at 60 degrees C. Complete protection was observed at 5 mM AT and 1 mM Cap. Comparison of AT and Cap in their effects on melting temperature and heat stress-induced degradation showed that a greater protection occurs with Cap which has a weaker effect on melting temperature. Based on this observation it was concluded that the observed protection by AT may be explained by its effects on melting temperature while that of Cap should be ascribed to other mechanisms.
Arakawa
T
T
Alliance Protein Laboratories, 3957 Corte Cancion, 91360, Thousand Oaks, CA 91360, USA. tarakawa2@aol.com
Kita
Y
Y
D000818:Animals; D002210:Caprylates / Q000737:chemistry*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry
D002210:Caprylates; D012710:Serum Albumin, Bovine; C006392:N-acetyltryptophan; D014364:Tryptophan; C031492:octanoic acid