Analysis of structural changes in putidaredoxin by UV-Vis Spectroscopy (CAT#: STEM-MB-0907-WXH)

Introduction

Iron – sulfur proteins are found in all organisms and function as electron transfer mediators in complex redox systems. Putidaredoxin (Pdx) is an 11,400-Da protein, which serves as an electron shuttle between the flavin of the putidaredoxin reductase (PdR) and a heme in the cytochrome P450cam (CYP101). Pdx sequentially transfers two electrons to the heme during the enzymatic cycle of the stereospecific camphor hydroxylation.

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Principle Applications Procedure Materials Notes Related Products

Principle

UV-Vis spectroscopy is an analytical technique that measures the amount of discrete wavelengths of UV or visible light that are absorbed by or transmitted through a sample in comparison to a reference or blank sample. This property is influenced by the sample composition, potentially providing information on what is in the sample and at what concentration. The only requirement is that the sample absorb in the UV-Vis region, i.e. be a chromophore. Absorption spectroscopy is complementary to fluorescence spectroscopy. Parameters of interest, besides the wavelength of measurement, are absorbance (A) or transmittance (%T) or reflectance (%R), and its change with time.

Applications

UV/Vis spectroscopy is routinely used in analytical chemistry for the quantitative determination of diverse analytes or sample, such as transition metal ions, highly conjugated organic compounds, and biological macromolecules. Spectroscopic analysis is commonly carried out in solutions but solids and gases may also be studied.

Procedure

1. Calibrate the Spectrometer
2. Perform an Absorbance Spectrum
3. Kinetics Experiments with UV-Vis Spectroscopy

Materials

UV/VIS Spectrophotometer

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