title: Structural changes of beta-lactoglobulin during thermal unfolding and refolding--an FT-IR and circular dichroism study.
abstract: We have quantitatively characterized by FT-IR spectroscopy the contents of secondary structure of beta-lactoglobulin during thermal unfolding and subsequent refolding. Our data clearly indicate that considerable amount of secondary structure, particularly beta-sheet, still remained intact even at 90 degrees C. Noticeable changes in secondary structure of beta-lactoglobulin were observed only above 70 degrees C. The refolded protein regained, within limits of experimental error, all of the secondary structure lost during thermal unfolding. The data also indicate that the refolding mechanism operating at pH 7.0 and 2.0 are the same. Identical secondary structure of native and refolded beta-lactoglobulin was also indicated by far-UV circular dichroic spectra of the two forms of protein. Near UV circular dichroic spectra of the same two forms showed considerable differences indicating less tertiary structure of refolded beta-lactoglobulin. The combined CD and FT-IR data indicated that refolded form of beta-lactoglobulin could be characterized as a molten globule state as it had native-like secondary structure and compromised tertiary structure.
authors.affiliation: Protein Chemistry Laboratory, Department of Chemistry, Kolkata 700 009, India.
authors.forename: C
authors.initials: C
authors.lastname: Bhattacharjee
authors.forename: S
authors.initials: S
authors.lastname: Saha
authors.forename: A
authors.initials: A
authors.lastname: Biswas
authors.forename: M
authors.initials: M
authors.lastname: Kundu
authors.forename: L
authors.initials: L
authors.lastname: Ghosh
authors.forename: K P
authors.initials: KP
authors.lastname: Das
mesh_terms: D002942:Circular Dichroism; D006358:Hot Temperature; D007782:Lactoglobulins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared; D013816:Thermodynamics
chemical_list: D007782:Lactoglobulins