Process for screening of a binding amphiphilic peptides specific for hairpin RNA
The present invention relates to a screening method of an amphiphilic peptide specifically binding to hairpin RNA, more precisely a screening method of an amphiphilic peptide having specificity and strong binding strength to target hairpin RNA using peptide library comprising those peptides having modifications of both hydrophilic face and hydrophobic face. The method of the present invention provides a screening method of an amphiphilic peptide which is specific to hairpin RNA. So, the peptide selected by the method of the present invention can be effectively used for the study of hairpin RNA functions and for the production of a novel drug using an artificial peptide binding to a hairpin RNA target.
1. A method of screening an amphiphilic peptide binding specifically to a target hairpin RNA for production of a diagnostic reagent for hepatitis C virus comprising the steps of:
1) constructing an amphiphilic peptide library containing the amphiphilic peptide consisting of the amino acid sequences of SEQ. ID. NO: 1-No: 37;
2) synthesizing Internal Ribosome Entry Site (IRES) RNA as the target hairpin RNA of hepatitis C virus;
3) calculating binding force between RNA and the amphiphilic peptide by measuring fluorescent anisotropy of a mixture composed of the amphiphilic peptide, hairpin RNA and a fluorescent compound by using a fluoro spectrophotometer; and
4) selecting the amphiphilic peptide, wherein said amphiphilic peptide has a dissociation constant less than 4.3 nM or a differentiation coefficient more than 4.9.
2. The method of screening according to claim 1 , wherein the compound is rhodamine conjugated rev peptide (Rev-N-rhodamine).
3. The method of screening according to claim 1 , wherein the selection of step 4) comprises selecting the amphiphilic peptide, wherein said amphiphilic peptide has a dissociation constant less than 0.87 nM or a differentiation coefficient more than 14.
4. The method of screening according to claim 1 , wherein the selection of step 4) comprises selecting the amphiphilic peptide, wherein said amphiphilic peptide has a dissociation constant less than 0.62 nM or a differentiation coefficient more than 28.