Pressure-Induced Secondary Structural Changes of Proteins Studied by FTIR Spectroscopy

Attempts were made to enhance the hydrolysis of Asclepias syriaca (As) seed floss and poplar seed floss (PSF) by cellulase after pre-treatment with ionic liquids. Two ionic liquids, namely 1-butyl-3-methylimidazolium chloride [BMIM]Cl and 1-ethyl-3-methylimidazolium tetrachloroaluminate [EMIM]Cl-AlCl3, were used. In comparison with conventional cellulose pretreatment processes, the ionic liquids were used under a milder condition corresponding to the optimum activity of cellulase. Hydrolysis kinetics of the IL-treated cellulose materials was significantly enhanced. The initial hydrolysis rates for IL-treated cellulose materials were greater than those of non-treated ones. The structural modifications of hydrolyzed cellulose materials were analyzed using FTIR spectroscopy.

Download Full-text

The application of ATR-FTIR Spectroscopy and the Reversible DNA Conformation as a Sensor to Test the Effectiveness of Platinum(II) Anticancer Drugs

10.20944/preprints201810.0406.v1 ◽

2018 ◽

Cited By ~ 1

Author(s):

Khansa Al-Jorani ◽

Anja Rüther ◽

Miguela Martin ◽

Rukshani Haputhanthri ◽

Glen B. Deacon ◽

...

Keyword(s):

Ftir Spectroscopy ◽

Cancer Cells ◽

Structural Changes ◽

Isotonic Saline ◽

Platinum Complexes ◽

Attenuated Total Reflection ◽

Dna Conformation ◽

Acetone Water ◽

Intensity Changes ◽

And Shifts

Platinum(II) complexes have been found to be effective against cancer cells. Cisplatin curbs cell replication by interacting with the deoxyribonucleic acid (DNA), eventually leading to cell death and reducing cell proliferation. In order to investigate the ability of platinum complexes to affect cancer cells, two examples from the class of polyflurophenylorganoamidoplatinum(II) complexes were synthesised and tested on isolated DNA. The two compounds trans-[N,N’-bis(1,2,3,5,6-pentafluorophenyl)ethane-1,2-diaminato(1-)](2,3,4,5,6-pentafluorobenzoato)(pyridine)platinum(II) (PFB), and trans-[N,N’-bis(1,2,3,5,6-pentafluorophenyl)ethane-1,2-diaminato(1-)](2,4,6-trimethylbenzoato)(pyridine)platinum(II) (TMB) were compared with cisplatin through their reaction with DNA. Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectroscopy was applied to analyse the interaction of the Pt(II) complexes with DNA in the hydrated, dehydrated and rehydrated state. These were compared with control DNA in acetone/water (PFB, TMB) and isotonic saline (cisplatin) under the same conditions. Principle Component Analysis (PCA) was applied to compare the ATR-FTIR spectra of the untreated control DNA with spectra of PFB and TMB treated DNA samples. Disruptions in the conformation of DNA treated with the Pt(II) complexes upon rehydration were mainly observed by monitoring the position of the IR-band around 1711 cm-1 assigned to the DNA base-stacking vibration. Furthermore, other intensity changes in the phosphodiester bands of DNA at ~1234 cm-1 and 1225 cm-1 and shifts in the dianionic phosphodiester vibration at 966 cm-1 were observed. The isolated double stranded DNA (dsDNA) or single stranded DNA (ssDNA) showed different structural changes when incubated with the studied compounds. PCA confirmed PFB had the most dramatic effect by denaturing both dsDNA and ssDNA. Both compounds, along with cisplatin, induced changes in DNA bands at 1711, 1088, 1051 and 966 cm-1 indicative of DNA conformation changes. The ability to monitor conformational change with infrared spectroscopy paves the way for a sensor to screen for new anticancer therapeutic agents.

Download Full-text

Structural analysis of proteins by isotope-edited FTIR spectroscopy

Spectroscopy An International Journal ◽

10.1155/2010/634831 ◽

2010 ◽

Vol 24 (1-2) ◽

pp. 37-43 ◽

Cited By ~ 11

Author(s):

Suren A. Tatulian

Keyword(s):

Structural Analysis ◽

Ftir Spectroscopy ◽

Structural Changes ◽

Isotope Labeling ◽

Structural Information ◽

Attenuated Total Reflection ◽

Site Specific ◽

Segmental Isotope Labeling ◽

Protein Membrane

Structure determination of multidomain proteins or protein–membrane complexes is one of the most challenging tasks in modern structural biology. High-resolution techniques, like NMR or X-ray crystallography, are limited to molecules of moderate size or those that can be crystallized easily. Both methods encounter serious technical obstacles in structural analysis of protein–membrane systems. This work describes an emerging biophysical technique that combines segmental isotope labeling of proteins with Fourier transform infrared (FTIR) spectroscopy, which provides site-specific structural information on proteins and allows structural characterization of protein–membrane complexes. Labeling of a segment of the protein with13C results in infrared spectral resolution of the labeled and unlabeled parts and thus allows identification of structural changes in specific domains/segments of the protein that accompany functional transitions. Segmental isotope labeling also allows determination of the precise configuration of protein–membrane complexes by polarized attenuated total reflection FTIR (ATR–FTIR) spectroscopy. These new developments offer solutions to functionally important site-specific structural changes in proteins and protein–membrane complexes that are hard to approach using conventional methods.

Download Full-text