Peer-Reviewed Journal Details
Mandatory Fields
Kaminska, A;Forster, RJ;Keyes, TE
2010
February
Journal of Raman Spectroscopy
The impact of adsorption of bovine pancreatic trypsin inhibitor on CTAB-protected gold nanoparticle arrays: a Raman spectroscopic comparison with solution denaturation
Published
7 ()
Optional Fields
PROTEIN-STRUCTURE BEHAVIOR SILVER REFINEMENT MONOLAYERS SCATTERING RESOLUTION STABILITY MOLECULES CRYSTAL
41
130
135
The influence of an array of cetyltrimethylammonium bromide (CTAB)-protected gold nanoparticles on the structure of a model protein, bovine pancreatic trypsin inhibitor (BPTI) at pH 7.4, was studied by Raman spectroscopy. The structural consequences of array adsorption were compared with the effects of deposition of the protein directly onto a roughened gold substrate and with thermal and reductive treatment of BPTI in solution. Both thermal and reductive denaturation in solution result in loss of alpha-helix structure, with an increase in random conformations of the protein in the case of reductive denaturation and beta-sheet conformation and random coil on thermal denaturation. For reductive denaturation in particular, extensive loss of secondary structure is evident. Deposition of the protein onto the array resulted in increased beta-sheet conformation similar to that observed on thermal treatment of the protein. However, unlike denaturation, which for both thermal and reductive process resulted in changes in the disulfide stretching wavenumber, this remains largely unchanged on application of the protein to the array. Furthermore, deposition of the protein onto bare gold results in significant heterogeneity in the S-S stretching signal with appearance of ggt and nonequilibrium geometry of the CCSSCC dihedral angles. Thermal denaturation results in a red shift of the SS mode, whereas dithiothreitol (DTT) treatment, as expected, induces significant loss of the S-S stretching signal, although a signal at 517 cm(-1) remains suggesting that the unreduced disulfide has changed to the ggt geometry. In addition, an SH mode is observed at 2570 cm(-1) in solution. The response of BPTI to thermal and DTT treatment while on the array is very different to its solution behavior, and suggests that adhesion to the array increases the stability of the protein. Copyright (C) 2009 John Wiley & Sons, Ltd.
CHICHESTER
0377-0486
10.1002/jrs.2477
Grant Details