Figure 2.

Glycerol gradient centrifugation of TelN protein. A 3.8-ml linear gradient from 15% to 35% (wt/vol) glycerol was overlaid with a TelN protein solution (Table 1, fraction V; 120 μl containing 400 μg of protein). Centrifugation was carried out at 100,000 × g for 15 h at 4°C. Fourteen fractions of 270 μl each were collected; 15 μl of each fraction and a 3-μl aliquot of the TelN protein loaded onto the gradient were separated by using a 15% SDS/PAGE gel and stained with Serva Blue R after electrophoresis. The fractions were assayed for TelN activity, as described in Materials and Methods. The amount of protein contained within each fraction and the TelN activity of each fraction (relative to the total amount and activity applied) are shown in the diagram (▴, relative enzymatic activity; ♦, relative amount of protein). The direction of sedimentation is right to left. Arrows indicate the positions of peaks of the reference proteins [1, catalase (11.3); 2, aldolase (7.8); and 3, BSA (4.4)]. The sedimentation coefficient is given in parenthesis in Svedberg units (S_20,W).