Use of a Polyethylene Glycol-Peptide Conjugate in a Competition Gel Shift Assay for Screening Potential Antagonists of HIV-1 Tat Protein Binding to TAR RNA

Jihong Wang, Shaei Yun Huang, Indrani Choudhury, Michael J Leibowitz, Stanley Stein

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Interference of binding of Tat protein to TAR RNA in HIV-1-infected cells may be a useful therapeutic strategy for AIDS. An electrophoretic assay to screen potential low-molecular-weight (<2 kDa) Tat antagonists has been established. A radiolabeled TAR RNA fragment (ΔTAR) is retarded in mobility when bound by a Tat peptide-polyethylene glycol conjugate (Tat-PEG), which is used in place of the Tat protein. The assay determines the ability of a potential antagonist to compete with Tat-PEG for binding to ΔTAR, as measured by interference with the gel shift of ΔTAR. To discriminate between specific and nonspecific interactions, the assay is done in the absence or the presence of a 250-fold molar excess of tRNA.

Original languageEnglish (US)
Pages (from-to)238-242
Number of pages5
JournalAnalytical Biochemistry
Volume232
Issue number2
DOIs
StatePublished - Dec 10 1995
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

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