title: A Distinct Proof on Interplay between Trehalose and Guanidinium Chloride for the Stability of Stem Bromelain.
abstract: Guanidinium chloride (GdnHCl), a potential denaturant, is well-known to denature a number of proteins in vitro as well as in vivo studies. Its deleterious action on stem bromelain (BM) is quite prominent resulting decrease in protein structure and stability. The counteraction of this adverse effect of GdnHCl by the use of osmolytes is scarcely studied and the mechanism is still illusive and not exclusive. For the first time, to test elegant and simple counteraction hypothesis as a general mechanism we utilized fluorescence, circular dichroism, Fourier transform infrared spectroscopy, and dynamic light scattering to study the counteraction of GdnHCl-induced denaturation of BM by the trehalose. It is revealed from the investigation of the results that trehalose is efficiently counteracting GdnHCl undesirable impacts on BM stability at molar ratio 1:1 of trehalose and GdnHCl. On the contrary, proteolytic activity of BM is increased only for the counteraction study of BM at very high concentrations of GdnHCl although still less than BM in buffer. The mutual exclusion of both trehalose and GdnHCl may stand for the counteraction of denaturation of BM resulting in a compact conformation with less solvent exposed surface area and increased secondary and tertiary structures. In addition, a decrease in BM-solvent interactions may also be contributing to some extent as there is little binding of trehalose replacing some water molecules and reducing binding of GdnHCl.
authors.affiliation: Department of Chemistry, University of Delhi , Delhi 110 007, India.
authors.forename: Anjeeta
authors.initials: A
authors.lastname: Rani
authors.affiliation: Department of Chemistry, University of Delhi , Delhi 110 007, India.
authors.forename: Pannuru
authors.initials: P
authors.lastname: Venkatesu
mesh_terms: D001963:Bromelains / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D000067493:Dynamic Light Scattering; D004795:Enzyme Stability / Q000187:drug effects; D019791:Guanidine / Q000737:chemistry / Q000494:pharmacology*; D011489:Protein Denaturation / Q000187:drug effects*; D017433:Protein Structure, Secondary / Q000187:drug effects; D058767:Protein Unfolding / Q000187:drug effects; D017550:Spectroscopy, Fourier Transform Infrared; D014199:Trehalose / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D001963:Bromelains; D014199:Trehalose; C091760:stem bromelain; D019791:Guanidine