FIG. 2.

Determination of the oligomerization of the peptides DCD-1L, LEK-45, SSL-23, and LL-37 in solution. (A) Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of human eccrine sweat and 4 μg of the DCD peptides LEK-45 and DCD-1L dissolved in water using a polyclonal anti-DCD antibody, which detects the C terminus of DCD-1L. Seen are SDS-stable dimers for LEK-45 and DCD-1L and higher oligomers in sweat. (B) Percent fluorescence recovery of FITC-labeled peptides LL-37 (□), DCD1L (▪), and SSL-23 (░⃞) in 1× PBS (pH 7.4) at different concentrations (0.0625 to 0.25 μM). Peptides were preincubated 2 h in PBS before proteinase K (10 μg/ml) treatment. Oligomerization of the peptides in solution was determined by fluorescence dequenching. (C) Determination of the time-kinetics of oligomerization: peptides (0.25 μM) were incubated for different time points (0 to 120 min) at room temperature in PBS before proteinase K treatment, and the percentage of fluorescence recovery was determined.