Selection of single-stranded DNA molecules that bind and inhibit human thrombin

• Published in: Nature (London) 1992-02, Vol.355 (6360), p.564-566
• Main Authors: Bock, Louis C, Griffin, Linda C, Latham, John A, Vermaas, Eric H, Toole, John J
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Base Sequence; Biological and medical sciences; Blood; Blood Coagulation - drug effects; Cloning, Molecular; Coagulation; Deoxyribonucleic acid; DNA; DNA, Single-Stranded - isolation & purification; Dose-Response Relationship, Drug; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Humanities and Social Sciences; Humans; Hydrolases; In Vitro Techniques; letter; Medical research; Molecular Sequence Data; multidisciplinary; Nucleic acids; Polymerase Chain Reaction; Science; Science (multidisciplinary); Sequence Homology, Nucleic Acid; Thrombin - antagonists & inhibitors; Thrombin Time
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/355564a0

APTAMERS 1 are double-stranded DNA or single-stranded RNA molecules that bind specific molecular targets. Large randomly generated populations can be enriched in aptamers by in vitro selection and polymerase chain reaction 1–11 . But so far single-stranded DNA has not been investigated for aptamer properties, nor has a target protein been considered that does not interact physiologically with nucleic acid. Here we describe the isolation of single-stranded DNA aptamers to the protease thrombin of the blood coagulation cascade and report binding affinities in the range 25–200 nM. Sequence data from 32 thrsombin aptamers, selected from a pool of DNA containing 60 nucleotides of random sequence, displayed a highly conserved 14–17–base region. Several of these aptamers at nanomolar concentrations inhibited thrombin-catalysed fibrin-clot formation in vitro using either purified fibrinogen or human plasma.