The fluorescence spectra were collected at 25°C using the PTI QuantaMaster 400 (Horiba, Canada) with monochromators for both excitation and emission. Fluorescence spectra of MMOH [0.32 μM MMOH in 300 μl of 25 mM Mops (pH 7.0) and 1 mM DTT] were determined using an excitation wavelength of 282 nm. Fluorescence was quantified by the integration of fluorescence emission bands with a maximum at 363 nm. The MMOH fluorescence was quenched by titration with MMOB, MMOD, and truncated MMODs (residues 1 to 74, 12 to 111, and 12 to 74). These proteins were titrated against MMOH in the concentration range from 0 to 15 μM. Dissociation constants were determined by fitting the intensity of emission at 363 nm (Origin 2018b and Wolfram Mathematica). Independent experiments were repeated in triplicate to allow the calculation of the average ± SEM.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.