Dynamics of human serum albumin studied by acoustic relaxation spectroscopy.
Sonic absorption spectra of solutions of human serum albumin (SA) in water and in aqueous phosphate buffer systems have been measured between 0.2 and 2000 MHz at different temperatures (15-35 degrees C), pH values (1.8-12.3), and protein concentrations (1-40 g/L). Several spectra, indicating relaxation processes in the whole frequency range, have been found. The spectra at neutral pH could be fitted well with an analytical function consisting of the asymptotic high frequency absorption and two relaxation contributions, a Debye-type relaxation term with discrete relaxation time and a term with asymmetric continuous distribution of relaxation times. Both relaxation contributions were observed in water and in buffer solutions and increased with protein concentration. The contribution represented by a Debye-type term is practically independent of temperature and was attributed to cooperative conformational changes of the polypeptide chain featuring a relaxation time of about 400 ns. The distribution of the relaxation times corresponding to the second relaxation contribution was characterized by a short time cutoff, between about 0.02 and 0.4 ns depending on temperature, and a long time tail extending to microseconds. Such relaxation behavior was interpreted in terms of solute-solvent interactions reflecting various hydration layers of HSA molecules. At acid and alkaline pH, an additional Debye-type contribution with relaxation time in the range of 30-100 ns exists. It seems to be due to proton transfer reactions of protein side-chain groups. The kinetic and thermodynamic parameters of these processes have been estimated from these first measurements to indicate the potential of acoustic spectra for the investigation of the elementary kinetics of albumin processes.
Hushcha
T
T
Institute of Bioorganic Chemistry and Petrochemistry, National Academy of Science of Ukraine, Murmans'ka Str. 1, 02094 Kiev, Ukraine. hushcha@bpci.kiev.ua
Kaatze
U
U
Peytcheva
A
A
D000162:Acoustics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D011522:Protons; D012709:Serum Albumin / Q000737:chemistry*; D013053:Spectrophotometry / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D010455:Peptides; D011522:Protons; D012709:Serum Albumin