title: Proteolytically-induced changes of secondary structural protein conformation of bovine serum albumin monitored by Fourier transform infrared (FT-IR) and UV-circular dichroism spectroscopy.
abstract: Enzymatically-induced degradation of bovine serum albumin (BSA) by serine proteases (trypsin and α-chymotrypsin) in various concentrations was monitored by means of Fourier transform infrared (FT-IR) and ultraviolet circular dichroism (UV-CD) spectroscopy. In this study, the applicability of both spectroscopies to monitor the proteolysis process in real time has been proven, by tracking the spectral changes together with secondary structure analysis of BSA as proteolysis proceeds. On the basis of the FTIR spectra and the changes in the amide I band region, we suggest the progression of proteolysis process via conversion of α-helices (1654 cm(-1)) into unordered structures and an increase in the concentration of free carboxylates (absorption of 1593 and 1402 cm(-1)). For the first time, the correlation between the degree of hydrolysis and the concentration of carboxylic groups measured by FTIR spectroscopy was revealed as well. The far UV-CD spectra together with their secondary structure analysis suggest that the α-helical content decreases concomitant with an increase in the unordered structure. Both spectroscopic techniques also demonstrate that there are similar but less spectral changes of BSA for the trypsin attack than for α-chymotrypsin although the substrate/enzyme ratio is taken the same.
authors.affiliation: Institute of Biophysics, Johann Wolfgang Goethe Universität, Max-von-Laue-Str.1, D-60438 Frankfurt am Main, Germany; Center for Drug Research & Development and Pharmacokinetic Applications (ARGEFAR), Ege University, Izmir, Turkey. Electronic address: gunnurgorucu@gmail.com.
authors.forename: Günnur
authors.initials: G
authors.lastname: Güler
authors.affiliation: A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, 28 ul. Vavilova, 119991 Moscow, Russia.
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Vorob'ev
authors.affiliation: Institute of Biophysics, Johann Wolfgang Goethe Universität, Max-von-Laue-Str.1, D-60438 Frankfurt am Main, Germany.
authors.forename: Vitali
authors.initials: V
authors.lastname: Vogel
authors.affiliation: Institute of Biophysics, Johann Wolfgang Goethe Universität, Max-von-Laue-Str.1, D-60438 Frankfurt am Main, Germany. Electronic address: maentele@biophysik.uni-frankfurt.de.
authors.forename: Werner
authors.initials: W
authors.lastname: Mäntele
mesh_terms: D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism; D002942:Circular Dichroism; D000072756:Protein Conformation, alpha-Helical; D059748:Proteolysis; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D014466:Ultraviolet Rays
chemical_list: D012710:Serum Albumin, Bovine; D002918:Chymotrypsin
keywords: CD spectroscopy; Enzyme; FTIR spectroscopy; Protein conformation; Protein secondary structure; Proteolysis