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Affinity Capillary Electrophoresis 317
0.0050
0.0025
A
Modif. AP-1
AP-1
scrambled AP-1
AP-1:
EKPLQNFTLCFR
Modif. AP-1:
E*KPLQNFTLCFR
Scrambled AP-1:
TRLFPKECLNQF
M
0.0000
A200 nm
-0.0025
0.0050
6 7 8 9 10 11 12
B
0.0025
0.0000
-0.0025
6 7 8 9 10 11 12
Time (min.)
+ Heparin
Fig. 4. Capillary electrophoresis (CE)-based binding study using synthetic SAP-
T3-derived peptides (c.f. Fig. 3) elucidate structure-function relationships of heparinbinding
peptides. Electrophoresis buffer was 0.1 M sodium phosphate, pH 7.4. The
separation took place in a 50-m inner diameter uncoated fused silica capillary with
50 cm to the detector window and of 57 cm total length. Separations were carried
out at 18 kV at liquid cooling at 20 C. Samples were pressure injected for 8 s after
a 2-s pre-injection of water and were subjected to electrophoresis from a separate
set of buffer vials than those used for pre-rinse. Peptide structures are indicated
using single-letter amino acid abbreviations. The AP-1 peptide preparation used for
the CE experiments was found to contain a mixture of regular AP-1 and modified
(dehydrated) AP-1 (modif. AP-1), while scrambled AP-1 was homogeneous. A 1:1
mixture of AP-1 peptide and the scrambled AP-1 peptide (both 0.5 mg/ml (334
M) in water) were analysed using CE in the absence (A) or presence (B) of 1
mg/ml (200 M) LMW heparin in the electrophoresis buffer (Adapted with permission
from (110)).