Esposito, Christina (2000) Investigation of the interaction of GdnHCl with poly-L-lysine, horseradish peroxidase and cytochrome c monitored via FTIR, CD and fluorescence spectroscopies. Masters thesis, Concordia University.
Spectroscopic probes were utilized to study the interaction of GdnHCl with three model systems: poly-L-lysine (PLL), horseradish peroxidase (HRP) and cytochrome c. Probing the chemically denatured states of proteins by FTIR is a novel technique, which has only recently been reported. To aid in the interpretation of the FTIR results, fluorescence and CD were employed. The use of deuterated solvents, i.e. Gdn-d 5 -DCl and D 2 O, simplified data processing and is recommended for FTIR studies of protein chemical denaturation. PLL, a random coil, was used to model the interaction between Gdn-d 5 -DCl and random structures. Spectroscopic evidence strongly suggested that Gdnd 5 -DCl associates with PLL, particularly with its lysyl side chains. The analysis of HRP indicated that it was not a suitable model due to complex time-dependent spectral changes. The study of cytochrome c in Gdn-d 5 -DCl revealed that the denaturant preferentially interacted with random structures. The denatured states of horse and tuna c were characterized and subtle differences between the two species were reported. Curvefitting of the FTIR amide I ' mode (1600-1700 cm -1 ) revealed that tuna c contained more residual structure in Gdn-d 5 -DCl than horse c. It was successfully demonstrated that FTIR is a powerful tool for probing the chemically denatured states of model protein systems, with potential applications for other biomolecules.
|Divisions:||Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry|
|Item Type:||Thesis (Masters)|
|Pagination:||xi, 137 leaves : ill. ; 29 cm.|
|Degree Name:||Theses (M.Sc.)|
|Program:||Chemistry and Biochemistry|
|Thesis Supervisor(s):||English, Ann M.|
|Deposited By:||Concordia University Libraries|
|Deposited On:||27 Aug 2009 17:16|
|Last Modified:||08 Dec 2010 15:18|
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