Combinatorial synthesis of cholesterol ester transfer protein-mRNA ligands and screening by nondenaturating gel-electrophoresis.
RNA, as one of the biomolecules with the most structural and functional diversity, is an attractive therapeutic target.(1) Employing combinatorial chemistry methods, small peptide ligands were found, which bind to a short RNA with important biological functions. A 23-nt RNA oligonucleotide from the cholesterol ester transfer protein mRNA was chosen as a molecular target.(2) A 27-nt RNA oligonucleotide from the human immunodeficiency virus type-1 (HIV-1) TAR RNA was used to control the binding specificity.(3) Tetrapeptide libraries, composed of the amino acids Lys, Tyr, Leu, Ile, and Arg, with and without C- and N-terminal lysines, were synthesized by a combination of combinatorial and divergent solid-phase synthesis. Gel-shift affinity screening was used to extract the peptides with the best RNA binding properties. The peptide Lys-Tyr-Lys-Leu-Tyr-Lys-Cys-NH(2) (1) showing micromolar affinity to its RNA target was characterized with circular dichroism (CD), ultra violet (UV) measurement, and (1)H NMR spectroscopy.
Baumann
M
M
Institut für Organische Chemie, Johann Wolfgang Goethe-Universität Frankfurt/Main, Marie-Curie-Str. 11, 60439 Frankfurt, Germany.
Bischoff
H
H
Schmidt
D
D
Griesinger
C
C
D020121:5' Untranslated Regions; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis*; D053480:Cholesterol Ester Transfer Proteins; D002942:Circular Dichroism; D020650:Combinatorial Chemistry Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D011489:Protein Denaturation; D012333:RNA, Messenger / Q000187:drug effects*; D013056:Spectrophotometry, Ultraviolet
D020121:5' Untranslated Regions; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D006023:Glycoproteins; D008024:Ligands; D012333:RNA, Messenger