Structural evolution during protein denaturation as induced by different methods.
Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) have been used to study conformational changes in protein bovine serum albumin (BSA) due to perturbation in its native structure as induced by varying temperature and pressure, and in presence of protein denaturating agents urea and surfactant. BSA has prolate ellipsoidal shape at ambient temperature and we observe no effect of temperature on its structure up to a temperature of about 60 degrees C . At temperatures beyond 60 degrees C , protein denaturation leads to aggregation. The protein solution exhibits a fractal structure at temperatures above 64 degrees C , and its fractal dimension increases with temperature. This is an indication of aggregation followed by gelation that evolves with increasing temperature. It is known for some of the proteins (e.g., Staphylococcal Nuclease) that pressure of 200 MPa can unfold the protein, whereas BSA does not show any protein unfolding even up to the pressure of 450 MPa . In presence of urea, the BSA protein unfolds for urea concentrations greater than 4M and acquires a random coil configuration. We make use of the dilution method to show the reversibility of protein unfolding with urea. The addition of surfactant denaturates the protein by the formation of micellelike aggregates of surfactants along the unfolded polypeptide chains of the protein. We show such structure of the protein-surfactant complex can be stabilized at higher temperatures, which is not the case for pure protein.
Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai-400 085, India.
S
S
Chodankar
V K
VK
Aswal
J
J
Kohlbrecher
R
R
Vavrin
A G
AG
Wagh
D000818:Animals; D001703:Biophysics / Q000379:methods*; D002417:Cattle; D017709:Fractals; D008027:Light; D008836:Micrococcal Nuclease / Q000737:chemistry; D015233:Models, Statistical; D008968:Molecular Conformation; D009502:Neutrons; D011489:Protein Denaturation; D012542:Scattering, Radiation; D012709:Serum Albumin / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry; D013696:Temperature
D012709:Serum Albumin; D013501:Surface-Active Agents; D008836:Micrococcal Nuclease