Han, KC Yu, JH Yang, EG 2024-01-21T04:09:18Z 2024-01-21T04:09:18Z 2021-09-02 2005-11 0173-0835 https://pubs.kist.re.kr/handle/201004/136037 Human immunodeficiency virus type 1 (HIV-1) Rev protein is known to regulate the expression of proteins via binding to an RNA site termed the HIV Rev response element (RRE) presumably with a defined shape, mediated mainly by electrostatic interactions. We have developed a quantitative method based on CE-LIF detection for a systematic evaluation of interactions between a truncated RRE (tRRE) RNA and an HIV-1 Rev peptide. Employing a fluorescently labeled HIV-1 Rev protein fragment (RevF) as a probe, buffers were evaluated for the separation and detection as well as for the RNA shape-specific formation of the complex. Selection of an optimal buffer condition allowed us to perform quantitation of the tRRE-RevF complex formation and determine its dissociation constant. In addition, competitive inhibitions of the RNA-peptide interaction by some aminoglycosides were evaluated quantitatively by monitoring the complex peak, resulting in determination Of IC50 values. This sensitive and reliable CE-LIF-based method would be of interest in developing various screening systems for English WILEY AFFINITY INTERACTIONS RNA TARGETS BINDING PROTEIN RRE AMINOGLYCOSIDES EXPRESSION INHIBITOR REGION Quantitative analysis of shape-specific interactions of Rev response element with a positively charged Rev peptide by capillary electrophoresis Article 10.1002/elps.200500154 1 ELECTROPHORESIS, v.26, no.22, pp.4379 - 4386 ELECTROPHORESIS 26 22 4379 4386 scie scopus 000233740900018 2-s2.0-28244492757 Biochemical Research Methods Chemistry, Analytical Biochemistry & Molecular Biology Chemistry Article AFFINITY INTERACTIONS RNA TARGETS BINDING PROTEIN RRE AMINOGLYCOSIDES EXPRESSION INHIBITOR REGION CE-LIF Rev peptide Rev response element Rev responsive element-Rev interaction