Understanding dynamics of myoglobin in heterogeneous aqueous environments using coupled water fractions.
This work presents an analysis of near environment of myoglobin (Mb) in different aqueous solutions (in the presence of NaCl, sucrose, trehalose, urea, and glycerol) using the coupled water fractions measured using a quartz crystal microbalance (QCM). The secondary structural features of the protein from circular dichroic (CD) spectroscopy and the coupled water fractions give important clues to the overall dynamics of the protein. Using time resolved fluorescence, these leads have been applied to understand the observed lifetime relaxations of Mb. Though the time scales of observation of coupled water and the lifetimes are very different, our study suggests that the trends in coupled water fraction seem to be good indicators for regulation of the relaxation dynamics of the protein. The relaxations generally show a triphasic distribution of time scales. The initial relaxation in the picoseconds time scale represents the local motions of coupled water followed by a slightly slower decay in hundreds of picoseconds attributable to coupled water-'quasi free' water interactions. The third nanosecond lifetime is due to changes in transitions in isomers of hydrated protein. The dynamics of coupled water in Mb with NaCl is the fastest (around 21 ps) and is slowest in glycerol (250 ps). The results strongly indicate that it is the resident times of water molecules that play a dominant role in the overall stability of protein in a particular hydrated isomer and not just always the number of such water molecules in the hydrated protein.
Muthuselvi
L
L
Chemical Lab., CLRI (CSIR), Adyar, Chennai 600 020, India.
Dhathathreyan
Aruna
A
D002021:Buffers; D002942:Circular Dichroism; D017666:Deuterium Oxide; D005990:Glycerol; D008958:Models, Molecular; D009211:Myoglobin / Q000737:chemistry*; D011487:Protein Conformation; D012965:Sodium Chloride; D012997:Solvents / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013395:Sucrose; D014199:Trehalose; D014508:Urea; D014867:Water / Q000737:chemistry*
D002021:Buffers; D009211:Myoglobin; D012997:Solvents; D014867:Water; D012965:Sodium Chloride; D013395:Sucrose; D014508:Urea; D014199:Trehalose; D017666:Deuterium Oxide; D005990:Glycerol