Solution Behavior and Interaction of Pepsin with Carnitine Based Cationic Surfactant: Fluorescence, Circular Dichroism, and Calorimetric Studies.
The present work reports the pH-induced conformational changes of pepsin in solution at room temperature. The conformational change makes the protein surface active. The protein was found to be present in the partially denatured state at pH 8 as well as at pH 2. The fluorescence probe and circular dichroism (CD) spectra suggested that the most stable state of pepsin exists at pH 5. The binding affinities of pepsin in its native and denatured states for a D,L-carnitine-based cationic surfactant (3-hexadecylcarbamoyl-2-hydroxypropyl)trimethylammonium chloride (C16-CAR) were examined at very low concentrations of the surfactant. The thermodynamics of the binding processes were investigated by use of isothermal titration calorimetry. The results were compared with those of (3-hexadecylcarbamoylpropyl)trimethylammonium chloride (C16-PTAC), which is structurally similar to C16-CAR, but without the secondary -OH functionality near the headgroup. None of the surfactants were observed to undergo binding with pepsin at pH 2, in which it exists in the acid-denatured state. However, both of the surfactants were found to spontaneously bind to the most stable state at pH 5, the partially denatured state at pH 8, and the alkaline denatured state at pH 11. Despite the difference in the headgroup structure, both of the surfactants bind to the same warfarin binding site. Interestingly, the driving force for binding of C16-CAR was found to be different from that of C16-PTC at pH ≥ 5. The steric interaction of the headgroup in C16-CAR was observed to have a significant effect on the binding process.
Department of Chemistry, Indian Institute of Technology Kharagpur , Kharagpur - 721 302, India.
Subhajit
S
Ghosh
Department of Chemistry, Indian Institute of Technology Kharagpur , Kharagpur - 721 302, India.
Subhrajyoti
S
Dolai
Department of Chemistry, Indian Institute of Technology Kharagpur , Kharagpur - 721 302, India.
Trilochan
T
Patra
Department of Chemistry, Indian Institute of Technology Kharagpur , Kharagpur - 721 302, India.
Joykrishna
J
Dey
D002151:Calorimetry / Q000379:methods*; D002331:Carnitine / Q000737:chemistry*; D002412:Cations; D002942:Circular Dichroism / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D010434:Pepsin A / Q000737:chemistry*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000737:chemistry*; D013816:Thermodynamics
D002412:Cations; D012996:Solutions; D013501:Surface-Active Agents; D010434:Pepsin A; D002331:Carnitine