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High-affinity aptamers selectively inhibit human nonpancreatic secretory phospholipase A2 (hnps-PLA2)
- Source :
- Journal of medicinal chemistry. 41(6)
- Publication Year :
- 1998
-
Abstract
- A family of sequence-related 2'-aminopyrimidine, 2'-hydroxylpurine aptamers, developed by oligonucleotide-based combinatorial chemistry, SELEX (systematic evolution of ligand by exponential enrichment) technology, binds human nonpancreatic secretory phospholipase A2 (hnps-PLA2) with nanomolar affinities and inhibits enzymatic activity. Aptamer 15, derived from the family, binds hnps-PLA2 with a Kd equal to 1.7 +/- 0.2 nM and, in a standard chromogenic assay of enzymatic activity, inhibits hnps-PLA2 with an IC50 of 4 nM, at a mole fraction of substrate concentration of 4 x 10(-6) and a calculated Ki of 0.14 nM. Aptamer 15 is selective for hnps-PLA2, having a 25- and 2500-fold lower affinity, respectively, for the unrelated proteins human neutrophil elastase and human IgG. Contractions of guinea pig lung pleural strips induced by hnps-PLA2 are abolished by 0.3 microM aptamer 15, whereas contractions induced by arachidonic acid are not altered. The structure that is essential for binding and inhibition appears to be a 40-base hairpin/loop motif with an asymmetrical internal loop. The affinity and activity of the aptamers demonstrate the ability of the SELEX process to isolate antagonists of nonnucleic-acid-binding proteins from vast oligonucleotide combinatorial libraries.
- Subjects :
- Male
Indoles
Aptamer
Guinea Pigs
Molecular Sequence Data
Oligonucleotides
In Vitro Techniques
Group II Phospholipases A2
Phospholipases A
chemistry.chemical_compound
Phospholipase A2
Drug Discovery
Animals
Humans
Amino Acid Sequence
Enzyme Inhibitors
IC50
Lung
Gene Library
biology
Base Sequence
Oligonucleotide
Elastase
Muscle, Smooth
Molecular biology
Phospholipases A2
chemistry
Biochemistry
Enzyme inhibitor
Drug Design
biology.protein
Molecular Medicine
Nucleic Acid Conformation
Pleura
RNA
lipids (amino acids, peptides, and proteins)
Arachidonic acid
Systematic evolution of ligands by exponential enrichment
Subjects
Details
- ISSN :
- 00222623
- Volume :
- 41
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Journal of medicinal chemistry
- Accession number :
- edsair.doi.dedup.....85cb4d0cd494e70739b0e4969db709a8