title: Effect of buffer species on the thermally induced aggregation of interferon-tau.
abstract: It is now becoming apparent that a common pathway of protein aggregation involves the unimolecular structural rearrangement from the native state to a slightly expanded aggregation-competent species. It is the goal of this study to understand the aggregation and the effects of buffer on the stability of IFN-tau. In this study, the thermally-induced aggregation of interferon-tau (IFN-tau) is described. By monitoring the aggregation rate in the presence of increasing amounts of sucrose, the relative change in surface area (Deltas) for conversion to the aggregation-competent state can be determined. Under conditions of pH 7 and in 20 mM buffer, the protein displays different aggregation rates depending on the nature of the buffer species. The protein aggregates mostly quickly in phosphate buffer, slower in the presence of Tris and slowest in the presence of histidine. The largest value for Deltas occurs for the histidine-containing samples, where aggregation proceeds via a slightly expanded aggregation competent state with a surface area increase of 7.6%. Furthermore, it appears that histidine binds to the native state of IFN-tau, thereby stabilizing the native state and retarding aggregation. Measurement of the second virial coefficient, B(22), for different formulations indicates that inclusion of histidine has only a small effect on repulsion between protein molecules, suggesting that colloidal stabilization is not the dominant mechanism for stabilization of IFN-tau. This study represents the first detailed biophysical study of specific buffer-induced stabilization, resulting in shifting the equilibrium towards the native state and away form the expanded aggregation-competent species.
authors.affiliation: Center for Pharmaceutical Biotechnology, University of Colorado Health Sciences Center, Denver, CO, USA.
authors.forename: Derrick S
authors.initials: DS
authors.lastname: Katayama
authors.forename: Rajiv
authors.initials: R
authors.lastname: Nayar
authors.forename: Danny K
authors.initials: DK
authors.lastname: Chou
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Valente
authors.forename: Julianne
authors.initials: J
authors.lastname: Cooper
authors.forename: Charles S
authors.initials: CS
authors.lastname: Henry
authors.forename: David G
authors.initials: DG
authors.lastname: Vander Velde
authors.forename: Lorelie
authors.initials: L
authors.lastname: Villarete
authors.forename: C P
authors.initials: CP
authors.lastname: Liu
authors.forename: Mark Cornell
authors.initials: MC
authors.lastname: Manning
mesh_terms: D000818:Animals; D002021:Buffers; D002151:Calorimetry; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004058:Diffusion; D004355:Drug Stability; D005470:Fluorometry; D006639:Histidine / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007370:Interferon Type I / Q000737:chemistry*; D007700:Kinetics; D008027:Light; D009994:Osmolar Concentration; D010710:Phosphates / Q000737:chemistry; D011257:Pregnancy Proteins / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D012542:Scattering, Radiation; D013816:Thermodynamics; D014325:Tromethamine / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D002021:Buffers; D007370:Interferon Type I; D010710:Phosphates; D011257:Pregnancy Proteins; C020947:interferon tau; D014325:Tromethamine; D014867:Water; D006639:Histidine