A DNA Spiegelmer to staphylococcal enterotoxin B.
Bacterial staphylococcal enterotoxin B is involved in several severe disease patterns and it was therefore used as a target for the generation of biologically stable mirror-image oligonucleotide ligands, so called Spiegelmers. The toxin is a 28 kDa protein consisting of 239 amino acids. Since the full-length protein is not accessible to chemical peptide synthesis, a stable domain of 25 amino acids was identified as a suitable selection target. DNA in vitro selection experiments were carried out against the equivalent mirror-image D-peptide domain resulting in high affinity D-DNA aptamers. As expected, the corresponding enantiomeric L-DNA Spiegelmer showed comparable binding characteristics to the L-peptide domain. Moreover, the Spiegelmer bound the whole protein target with only slightly reduced affinity. Dissociation constants of both peptide-oligonucleotide complexes were measured in the range of 200 nM, whereas the Spiegelmer binding to the full-length protein was determined at approximately 420 nM. These data demonstrate the possibility to identify Spiegelmers against large protein targets by a domain approach.
Purschke
Werner G
WG
NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
Radtke
Falko
F
Kleinjung
Frank
F
Klussmann
Sven
S
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004768:Enterotoxins / Q000737:chemistry* / Q000378:metabolism*; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D017434:Protein Structure, Tertiary
D004768:Enterotoxins; D008024:Ligands; D009838:Oligodeoxyribonucleotides; D010455:Peptides; C031818:enterotoxin B, staphylococcal; D004247:DNA