Analysis of Tertiary Interactions between SART3 and U6 Small Nuclear RNA Using Modified Nanocapillaries

Choongman Lee, Joon Kyu Park, Yeoan Youn, Joo Hyoung Kim, Kyo Seok Lee, Nak Kyoon Kim, Eunji Kim, Eunice Eunkyeong Kim, Kyung Hwa Yoo

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We employed modified glass nanocapillaries to investigate interactions between the RNA-binding protein, known as cell carcinoma antigen recognized by T cells-3 (SART3), and the noncoding spliceosome component, U6 small nuclear RNA (snRNA), at the single-molecule level. We functionalized the nanocapillaries with U6 snRNA fragments, which were hybridized to DNA molecules and then covalently attached to the nanocapillary surface. When transported through the modified nanocapillaries, two different SART3-derived constructs, HAT-RRM1-RRM2 and RRM1-RRM2, exhibited resistive ionic current pulses with different dwell times, which represented their different binding affinities to tethered U6 snRNAs. The dissociation constants (KD), estimated from the bias voltage dependence of translocation events, were approximately 1.9 μM and 201 μM for HAT-RRM1-RRM2 and RRM1-RRM2, respectively. These values were comparable to corresponding values obtained with isothermal titration calorimetry, demonstrating that the modified glass nanocapillaries are applicable to analyses of protein-ligand interactions at the single-molecule level.

Original languageEnglish
Pages (from-to)2390-2397
Number of pages8
JournalAnalytical Chemistry
Volume89
Issue number4
DOIs
Publication statusPublished - 2017 Feb 21

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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