Physicochemical properties of thiol proteinase inhibitor isolated from goat pancreas.
Thiol proteinase inhibitors are crucial to proper functioning of all living tissues consequent to their cathepsin regulatory and myriad important biologic properties. Equilibrium denaturation of dimeric goat pancreas thiol proteinase inhibitor (PTPI), a cystatin superfamily variant has been studied by monitoring changes in the protein's spectroscopic and functional characteristics. Denaturation of PTPI in guanidine hydrochloride and urea resulted in altered intrinsic fluorescence emission spectrum, diminished negative circular dichroism, and loss of its papain inhibitory potential. Native like spectroscopic properties and inhibitory activity are only partially restored when denaturant is diluted from guanidine hydrochloride unfolded samples demonstrating that process is partially reversible. Coincidence of transition curves and dependence of transition midpoint (3.2M) on protein concentration in guanidine hydrochloride-induced denaturation are consistent with a two-state model involving a native like dimer and denatured monomer. On the contrary, urea-induced unfolding of PTPI is a multiphasic process with indiscernible intermediates. The studies demonstrate that functional conformation and stability are governed by both ionic and hydrophobic interactions.
Department of Biochemistry, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Medha
M
Priyadarshini
Rizwan H
RH
Khan
Bilqees
B
Bano
D000817:Anilino Naphthalenesulfonates; D000818:Animals; D055598:Chemical Phenomena; D002942:Circular Dichroism; D015891:Cystatins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019281:Dimerization; D005456:Fluorescent Dyes; D006041:Goats; D019791:Guanidine; D066298:In Vitro Techniques; D010179:Pancreas / Q000378:metabolism*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D014508:Urea
D000817:Anilino Naphthalenesulfonates; D015891:Cystatins; D005456:Fluorescent Dyes; C027132:1-anilino-8-naphthalenesulfonate; D014508:Urea; D019791:Guanidine