Targeted PEG-based bioconjugates enhance the cellular uptake and transport of a HIV-1 TAT nonapeptide

Srinivasan Ramanathan, Bo Qiu, Shahriar Pooyan, Guobao Zhang, Stanley Stein, Michael J. Leibowitz, Patrick J. Sinko

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38 Scopus citations


We previously described the enhanced cell uptake and transport of R.I-K(biotin)-Tat9, a large (∼1500 Da) peptidic inhibitor of HIV-1 Tat protein, via SMVT, the intestinal biotin transporter. The aim of the present study was to investigate the feasibility of targeting biotinylated PEG-based conjugates to SMVT in order to enhance cell uptake and transport of Tat9. The 29 kDa peptide-loaded bioconjugate (PEG:(R.I-Cys-K(biotin)-Tat9)8) used in these studies contained eight copies of R.I-K(biotin)-Tat9 appended to PEG by means of a cysteine linkage. The absorptive transport of biotin-PEG-3400 (0.6-100 μM) and the bioconjugate (0.1-30 μM) was studied using Caco-2 cell monolayers. Inhibition of biotin-PEG-3400 by positive controls (biotin, biocytin, and desthiobiotin) was also determined. Uptake of these two compounds was also determined in CHO cells transfected with human SMVT (CHO/hSMVT) and control cells (CHO/pSPORT) over the concentration ranges of 0.05-12.5 μM and 0.003-30 μM, respectively. Nonbiotinylated forms of these two compounds, PEG-3350 and PEG:(R.I-Cys-K-Tat9)8, were used in the control studies. Biotin-PEG-3400 transport was found to be concentration-dependent and saturable in Caco-2 cells (Km=6.61 μM) and CHO/hSMVT cells (Km=1.26 μM). Transport/uptake was significantly inhibited by positive control substrates of SMVT. PEG:(R.I-Cys-K(biotin)Tat9)8 also showed saturable transport kinetics in Caco-2 cells (Km=6.13 μM) and CHO/hSMVT cells (Km=8.19 μM). Maximal uptake in molar equivalents of R.I-Cys-K(biotin)Tat9 was 5.7 times greater using the conjugate versus the biotinylated peptide alone. Transport of the nonbiotinylated forms was significantly lower (P<0.001) in all cases. The present results demonstrate that biotin-PEG-3400 and PEG:(R.I-Cys-K(biotin)Tat9)8 interact with human SMVT to enhance the cellular uptake and transport of these larger molecules and that targeted bioconjugates may have potential for enhancing the cellular uptake and transport of small peptide therapeutic agents.

Original languageEnglish (US)
Pages (from-to)199-212
Number of pages14
JournalJournal of Controlled Release
Issue number3
StatePublished - Dec 13 2001

All Science Journal Classification (ASJC) codes

  • Pharmaceutical Science


  • Caco-2
  • Drug targeting
  • PEG conjugates
  • Peptide delivery
  • SMVT
  • Tat


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