Mechanism for stabilization of the molten globule state of papain by sodium n-alkyl sulfates: spectroscopic and calorimetric approaches.
Papain exists in a molten globule (MG) state at pH 2 and in this state protein tends to aggregate in the presence of lower concentrations of guanidine hydrochloride (GuHCl). Such aggregation is prevented if low concentrations of sodium n-alkyl sulfates are also present in the buffer; in addition, stabilization of the protein is also induced. The guanidine hydrochloride and temperature-induced unfolding of papain, in the presence of n-alkyl sulfates, indicate stabilization of the protein as seen from the higher transition midpoints when monitored by fluorescence, circular dichroism, and differential scanning calorimetry. However, a similar phenomenon is not seen under neutral conditions in the presence of n-alkyl sulfate concentrations. The effect of n-alkyl sulfates on the structure of the MG state of papain was utilized to investigate the contribution of hydrophobic interaction to the stability of the MG state. The Td values of the MG states of papain in the presence of n-alkyl sulfates at different concentrations showed substantial variation. The enhancement of Td values at the stability criterion of MG states corresponded with increasing chain length of the cited n-alkyl sulfates. The present results suggest that the hydrophobic interactions play important roles in stabilizing and preventing the aggregation of the MG state of papain.
Chamani
J
J
Department of Biology, Faculty of Science, Islamic Azad University-Mashhad Branch, Mashhad, Iran. chamani@ibb.ut.ac.ir
Heshmati
M
M
D002152:Calorimetry, Differential Scanning / Q000379:methods; D004355:Drug Stability; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D010206:Papain / Q000737:chemistry*; D017510:Protein Folding; D012964:Sodium / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013463:Sulfuric Acid Esters / Q000737:chemistry*; D013816:Thermodynamics
D013463:Sulfuric Acid Esters; D012964:Sodium; D010206:Papain