An RNA aptamer that recognizes a specific conformation of the protein calsenilin.
The generation of molecules that selectively recognize specific conformations of a protein is an important component of the elucidation protein function. We have used SELEX (Systematic Evolution of Ligands by EXponential enrichment) technology to produce aptamers that bind in a conformationally selective manner to calsenilin, which involved in Ca(2+)-mediated apoptotic signaling. Since the conformations of calsenilin are quite different in the presence and absence of Ca(2+), aptamers were selected against the dimeric protein both under calcium-bound and calcium-free conditions. We have found that aptamer-12 selectively binds to the dimeric form of the protein in the presence of calcium ion, while the binding of aptamer-2 does not discriminate between the Ca(2+) bound and unbound protein. Data obtained from biochemical and biophysical experiments suggest that a dominant conformation of calcium-bound calsenilin exists in one dominant conformation and that one aptamer can be generated to recognize this conformation. In addition, observation made in this effort that aptamers selected against the two different conformations of calsenilin have different characteristics suggest that aptamers can serve as a plausible tool for recognizing various conformations of proteins, even those caused by interactions with small molecules or ions such as Ca(2+).
Lee
Kyung Hyun
KH
Department of Chemistry and Education, Seoul National University, Seoul 151-742, Republic of Korea.
Jeong
Sunjoo
S
Yang
Eun Gyung
EG
Park
Yong-Keun
YK
Yu
Jaehoon
J
D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology*; D019281:Dimerization; D051605:Kv Channel-Interacting Proteins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation
D052157:Aptamers, Nucleotide; D051605:Kv Channel-Interacting Proteins; D002118:Calcium