Screening of RNA molecules inhibiting human acetylcholinesterase by virtue of systematic evolution of ligands by exponential enrichment.
OBJECTIVE
To acquire the specific RNA aptamers inhibiting human red blood cell (RBC) acetylcholinesterase (AChE).


METHODS
Systematic evolution of ligands by exponential enrichment (SELEX) aptamer against human red blood cell membrane AChE was selected by microtiter plate method in vitro. The specifity binding to AChE was determined by gel mobility shift analysis. Microcolorispectrophotometric method was used to measure the activity of AChE.


RESULTS
The aptamers to human RBC AChE were identified by 9 reiterative rounds. At the same concentration (2.25 micromol/L), the aptamers did not bind to the recombinant human butyrylcholinesterase (rhBChE) but specifically bound to human RBC AChE and inhibited the enzyme activity.


CONCLUSIONS
It is an effective way to isolate the specific AChE inhibitor from the vast oligonucleotide combinatorial library by virtue of SELEX.
Zhang
Xing-Mei
XM
Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.
Shao
Ning-Sheng
NS
Chi
Mu-Gen
MG
Sun
Man-Ji
MJ
D000110:Acetylcholinesterase / Q000378:metabolism*; D001665:Binding Sites; D002800:Cholinesterase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D020650:Combinatorial Chemistry Techniques; D004910:Erythrocyte Membrane / Q000737:chemistry; D006801:Humans; D008024:Ligands; D012313:RNA / Q000302:isolation & purification* / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000378:metabolism
D002800:Cholinesterase Inhibitors; D008024:Ligands; D016601:RNA-Binding Proteins; D012313:RNA; D000110:Acetylcholinesterase