In vitro selection of single-stranded DNA aptamers that bind human pro-urokinase.
Single-chain pro-urokinase is an inactive proenzyme form of human urokinase (urinary plasminogen activator) with a Mr of 50,000 which is converted to the active two-chain form by catalytic amounts of plasmin. It is used for thrombolytic therapy of acute myocardial infarction and acute ischemic stroke. We have isolated single-stranded DNA molecules with significantly increased binding affinity for human pro-urokinase by SELEX (systematic evolution of ligands by exponential enrichment) procedure from a pool of 10(15) molecules containing 24 randomized positions which are flanked by defined regions. ssDNA from this library was hybridized with helper "fixture", thus allowing the central random chain to fold into complex three-dimentional shapes. Sequencing data from pro-urokinase aptamers obtained after 12 selection cycles displayed a highly conserved 12-14 base region.
Skrypina
Natalia A
NA
Russian Cardiology Research and Development Center, Moscow, Russia. nas@cardio.ru
Savochkina
Larissa P
LP
Beabealashvilli
Robert Sh
RSh
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D014568:Urokinase-Type Plasminogen Activator / Q000737:chemistry / Q000378:metabolism*
D004277:DNA, Single-Stranded; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase