title: Thermodynamic analysis of three state denaturation of Peanut Agglutinin.
abstract: Peanut Agglutinin (PNA) is a homotetrameric protein with a very unusual open quaternary structure. During denaturation, it first dissociates into a molten globule like state, which subsequently undergoes complete denaturation. Urea denaturation of PNA at neutral pH has been studied by intrinsic fluorescence spectroscopy and has been fitted to a three state model, A4 <=> 4I <=> 4U, to get all the relevant thermodynamic parameters. Urea denaturation leads to continuous red shift of wavelength maxima. The molten globule like state is formed in a short range of urea concentration. Refolding of the denatured PNA has been attempted by intrinsic fluorescence study. Refolding by instantaneous dilution shows the occurrence of the formation of an intermediate at a relatively rapid rate, within few seconds. The transition from PNA tetramer to molten globule like state is found to have a DeltaG value of approximately 33 kcal/mole while it is approximately 8 kcal/mole for the transition from molten globule like state to a completely denatured state. This in turn indicates that the tetramerization in PNA contributes significantly to the stability of the oligomer.
authors.affiliation: Molecular Biophysics Unit, Indian Institute of Science, Bangalore, Karnataka, India.
authors.forename: Sagarika
authors.initials: S
authors.lastname: Dev
authors.forename: Nirmala Devi
authors.initials: ND
authors.lastname: K
authors.forename: Sharmistha
authors.initials: S
authors.lastname: Sinha
authors.forename: Avadhesha
authors.initials: A
authors.lastname: Surolia
mesh_terms: D004305:Dose-Response Relationship, Drug; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D008958:Models, Molecular; D015233:Models, Statistical; D019887:Peanut Agglutinin / Q000737:chemistry*; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry; D014508:Urea / Q000737:chemistry
chemical_list: D019887:Peanut Agglutinin; D014364:Tryptophan; D014508:Urea