Abstract
Controlling the activity of a protein is necessary for defining its function in vivo. RNA aptamers are capable of inhibiting proteins with high affinity and specificity, but this effect is not readily reversible. We describe a general method for discovering aptamers that bind and inhibit their target protein, but addition of a specific small molecule disrupts the protein-RNA complex. A SELEX protocol was used to raise RNA aptamers to the DNA repair enzyme, formamidopyrimidine glycosylase (Fpg), and neomycin was employed in each round to dissociate Fpg-bound RNAs. We identified an RNA molecule able to completely inhibit Fpg at 100 nM concentration. Importantly, Fpg activity is recovered by the addition of neomycin. We envision these ligand-regulated aptamers (LIRAs) as valuable tools in the study of biological phenomena in which the timing of molecular events is critical.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 907-913 |
| Number of pages | 7 |
| Journal | Chemistry and Biology |
| Volume | 9 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 2002 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Organic Chemistry
Cite this
Controlling protein activity with ligand-regulated RNA aptamers. / Vuyisich, Momchilo; Beal, Peter A.
In: Chemistry and Biology, Vol. 9, No. 8, 08.2002, p. 907-913.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Controlling protein activity with ligand-regulated RNA aptamers
AU - Vuyisich, Momchilo
AU - Beal, Peter A.
PY - 2002/8
Y1 - 2002/8
N2 - Controlling the activity of a protein is necessary for defining its function in vivo. RNA aptamers are capable of inhibiting proteins with high affinity and specificity, but this effect is not readily reversible. We describe a general method for discovering aptamers that bind and inhibit their target protein, but addition of a specific small molecule disrupts the protein-RNA complex. A SELEX protocol was used to raise RNA aptamers to the DNA repair enzyme, formamidopyrimidine glycosylase (Fpg), and neomycin was employed in each round to dissociate Fpg-bound RNAs. We identified an RNA molecule able to completely inhibit Fpg at 100 nM concentration. Importantly, Fpg activity is recovered by the addition of neomycin. We envision these ligand-regulated aptamers (LIRAs) as valuable tools in the study of biological phenomena in which the timing of molecular events is critical.
AB - Controlling the activity of a protein is necessary for defining its function in vivo. RNA aptamers are capable of inhibiting proteins with high affinity and specificity, but this effect is not readily reversible. We describe a general method for discovering aptamers that bind and inhibit their target protein, but addition of a specific small molecule disrupts the protein-RNA complex. A SELEX protocol was used to raise RNA aptamers to the DNA repair enzyme, formamidopyrimidine glycosylase (Fpg), and neomycin was employed in each round to dissociate Fpg-bound RNAs. We identified an RNA molecule able to completely inhibit Fpg at 100 nM concentration. Importantly, Fpg activity is recovered by the addition of neomycin. We envision these ligand-regulated aptamers (LIRAs) as valuable tools in the study of biological phenomena in which the timing of molecular events is critical.
UR - http://www.scopus.com/inward/record.url?scp=0036669759&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036669759&partnerID=8YFLogxK
U2 - 10.1016/S1074-5521(02)00185-0
DO - 10.1016/S1074-5521(02)00185-0
M3 - Article
C2 - 12204690
AN - SCOPUS:0036669759
VL - 9
SP - 907
EP - 913
JO - Cell Chemical Biology
JF - Cell Chemical Biology
SN - 2451-9448
IS - 8
ER -