Sedimentation velocity experiments were carried out at 42,000 rpm and 20°C in a ProteomeLab XL-L analytical ultracentrifuge, using 12-mm aluminum-Epon double-sector centerpieces in an An55Ti rotor. Protein concentrations were recorded in continuous mode using absorbance at 280 nm and interferometry. P MD variants were studied at three different concentrations, namely, 0.8, 1.3, and 5 mg/ml. The partial specific volume of wt P MD, solvent density, and viscosity, as calculated with SEDNTER (75), were 0.741 ml/g according to P MD structure (PDB code 3ZDO), 0.99821 g/cm3, and 0.01002 poise, respectively. The partial specific volume of MD variants was assumed to be identical to that of wt MD. The data recorded from moving boundaries were analyzed in terms of both discrete species and continuous size distribution function of sedimentation coefficient C(S) using the program SEDFIT (76).

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

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.