Effect of ionic liquid on the native and denatured state of a protein covalently attached to a probe: solvation dynamics study.
Effect of a room temperature ionic liquid (RTIL, [pmim][Br]) on the solvation dynamics of a probe covalently attached to a protein (human serum albumin (HSA)) has been studied using femtosecond up-conversion. For this study, a solvation probe, 7-diethylamino-3-(4-maleimidophenyl)-4-methylcoumarin (CPM) has been covalently attached to the lone cysteine group (cys-34) of the protein HSA. Addition of 1.5 M RTIL or 6 M GdnHCl causes a red shift of the emission maxima of CPM bound to HSA by 3 nm and 12 nm, respectively. The average solvation time 〈τ(s)〉 decreases from 650 ps (in native HSA) to 260 ps (~2.5 times) in the presence of 1.5 M RTIL and to 60 ps (~11 times) in the presence of 6 M GdnHCl. This is ascribed to unfolding of the protein by RTIL or GdnHCl and therefore making the probe CPM more exposed. When 1.5 M RTIL is added to the protein denatured by 6 M GdnHCl in advance, a further ~5 nm red shift along with further ~2 fold faster solvent relaxation (<τ> ~30 ps) is observed. Our previous fluorescence correlation spectroscopy study [D. K. Sasmal, T. Mondal, S. Sen Mojumdar, A. Choudhury, R. Banerjee, and K. Bhattacharyya, J. Phys. Chem. B 115, 13075 (2011)] suggests that addition of RTIL to the protein denatured by 6 M GdnHCl causes a reduction in hydrodynamic radius (r(h)). It is demonstrated that in the presence of RTIL and GdnHCl, though the protein is structurally more compact, the local environment of CPM is very different from that in the native state.
Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata 700032, India.
Rajdeep
R
Chowdhury
Supratik
S
Sen Mojumdar
Shyamtanu
S
Chattoraj
Kankan
K
Bhattacharyya
D016880:Anisotropy; D003374:Coumarins; D003545:Cysteine / Q000737:chemistry; D019791:Guanidine; D006801:Humans; D052578:Ionic Liquids / Q000737:chemistry*; D011487:Protein Conformation; D012709:Serum Albumin / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D003374:Coumarins; D052578:Ionic Liquids; D012709:Serum Albumin; D012996:Solutions; C030860:N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl)maleimide; D019791:Guanidine; D003545:Cysteine