Skip to main content

Main menu

  • Home
  • About the Journal
    • General Information
    • Scope
    • Editorial Board
    • Impact & Metrics
    • Benefits of Publishing
    • Advertising/Sponsorship
    • About the Biochemical Society
  • Current Issue
  • For Authors
    • Submit Your Paper
    • Submission Guidelines
    • Editorial Policy
    • Open Access Policy
    • Rights and Permissions
    • Biochemical Society Member Benefits
  • For Librarians
    • Subscriptions and Pricing
    • Check Your Usage
    • Terms and Conditions
      • Biochemical Journal- Terms and Conditions of Usage
    • Open Access Policy
    • FAQs for Librarians
    • Register for Free Trial
  • For Readers
    • Subscribe
    • Rights and Permissions
    • Biochemical Society Member Benefits
    • Journal Access for Biochemical Society Members
    • Request a Free Trial
  • Collections
    • Article Collections
    • Classic Articles
  • Help
    • Technical Support
    • Contact Us
  • Other Publications
    • NEW: Emerging Topics in Life Sciences
    • NEW: Neuronal Signaling
    • Clinical Science
    • Biochemical Journal
    • Biochemical Society Transactions
    • Bioscience Reports
    • Essays in Biochemistry
    • Biochemical Society Symposia
    • Cell Signalling Biology
    • Glossary of Biochemistry and Molecular Biology
    • The Biochemist
    • Biochemical Society

User menu

  • Log-in
  • Subscribe
  • Contact Us

Search

  • Advanced search
  • Other Publications
    • NEW: Emerging Topics in Life Sciences
    • NEW: Neuronal Signaling
    • Clinical Science
    • Biochemical Journal
    • Biochemical Society Transactions
    • Bioscience Reports
    • Essays in Biochemistry
    • Biochemical Society Symposia
    • Cell Signalling Biology
    • Glossary of Biochemistry and Molecular Biology
    • The Biochemist
    • Biochemical Society

Log-in

Sign-up for alerts  
  • My Cart
Biochemical Journal
Browse Archive
Advanced Search
  • Home
  • About the Journal
    • General Information
    • Scope
    • Editorial Board
    • Impact & Metrics
    • Benefits of Publishing
    • Advertising/Sponsorship
    • About the Biochemical Society
  • Current Issue
  • For Authors
    • Submit Your Paper
    • Submission Guidelines
    • Editorial Policy
    • Open Access Policy
    • Rights and Permissions
    • Biochemical Society Member Benefits
  • For Librarians
    • Subscriptions and Pricing
    • Check Your Usage
    • Terms and Conditions
    • Open Access Policy
    • FAQs for Librarians
    • Register for Free Trial
  • For Readers
    • Subscribe
    • Rights and Permissions
    • Biochemical Society Member Benefits
    • Journal Access for Biochemical Society Members
    • Request a Free Trial
  • Collections
    • Article Collections
    • Classic Articles
  • Help
    • Technical Support
    • Contact Us

Research article

Temperature-, concentration- and cholesterol-dependent translocation of L- and D-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells

Marjan M. Fretz, Neal A. Penning, Saly Al-Taei, Shiroh Futaki, Toshihide Takeuchi, Ikuhiko Nakase, Gert Storm, Arwyn T. Jones
Biochemical Journal Mar 26, 2007, 403 (2) 335-342; DOI: 10.1042/BJ20061808
Marjan M. Fretz
Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, Wales, U.K.Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, PO Box 80082, 3508 TB Utrecht, The Netherlands
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Neal A. Penning
Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, Wales, U.K.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Saly Al-Taei
Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, Wales, U.K.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Shiroh Futaki
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, JapanPRESTO, Japan Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Toshihide Takeuchi
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Ikuhiko Nakase
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Gert Storm
Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, PO Box 80082, 3508 TB Utrecht, The Netherlands
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Arwyn T. Jones
Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, Wales, U.K.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
  • For correspondence: jonesat@cardiff.ac.uk
  • Article
  • Figures
  • Info & Metrics
  • Supplementary Data
  • PDF
Loading

Abstract

Delineating the mechanisms by which cell-penetrating peptides, such as HIV-Tat peptide, oligoarginines and penetratin, gain access to cells has recently received intense scrutiny. Heightened interest in these entities stems from their ability to enhance cellular delivery of associated macromolecules, such as genes and proteins, suggesting that they may have widespread applications as drug-delivery vectors. Proposed uptake mechanisms include energy-independent plasma membrane translocation and energy-dependent vesicular uptake and internalization through endocytic pathways. In the present study, we investigated the effects of temperature, peptide concentration and plasma membrane cholesterol levels on the uptake of a model cell-penetrating peptide, L-octa-arginine (L-R8) and its D-enantiomer (D-R8) in CD34+ leukaemia cells. We found that, at 4–12 °C, L-R8 uniformly labels the cytoplasm and nucleus, but in cells incubated with D-R8 there is additional labelling of the nucleolus which is still prominent at 30 °C incubations. At temperatures between 12 and 30 °C, the peptides are also localized to endocytic vesicles which consequently appear as the only labelled structures in cells incubated at 37 °C. Small increases in the extracellular peptide concentration in 37 °C incubations result in a dramatic increase in the fraction of the peptide that is localized to the cytosol and promoted the binding of D-R8 to the nucleolus. Enhanced labelling of the cytosol, nucleus and nucleolus was also achieved by extraction of plasma membrane cholesterol with methyl-β-cyclodextrin. The data argue for two, temperature-dependent, uptake mechanism for these peptides and for the existence of a threshold concentration for endocytic uptake that when exceeded promotes direct translocation across the plasma membrane.

  • cell-penetrating peptide
  • endocytosis
  • fluorescence microscopy
  • KG1a cell line
  • leukaemia
  • octa-arginine

Abbreviations: CM, complete RPMI 1640 medium; CPP, cell-penetrating peptide; MβCD, methyl-β-cyclodextrin; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; PI, propidium iodide; R8, octa-arginine; SFM, serum-free RPMI 1640 medium; Tf, transferrin

  • The Authors Journal compilation, The Biochemical Society, London
View Full Text

Log in using your username and password

Enter your Biochemical Journal username.
Enter the password that accompanies your username.
Forgot your user name or password?

Log in through your institution

You may be able to gain access using your login credentials for your institution. Contact your library if you do not have a username and password.
If your organization uses OpenAthens, you can log in using your OpenAthens username and password. To check if your institution is supported, please see this list. Contact your library for more details.
Previous ArticleNext Article
Back to top

 

 

April 2007

Volume: 403 Issue: 2

Biochemical Journal: 403 (2)
  • Table of Contents
  • Table of Contents (PDF)
  • Index by author
  • Ed Board (PDF)
  • Front Matter (PDF)

Actions

Email

Thank you for your interest in spreading the word about Biochemical Journal.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Temperature-, concentration- and cholesterol-dependent translocation of L- and D-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells
(Your Name) has forwarded a page to you from Biochemical Journal
(Your Name) thought you would like to see this page from the Biochemical Journal web site.
Share
Temperature-, concentration- and cholesterol-dependent translocation of L- and D-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells
Marjan M. Fretz, Neal A. Penning, Saly Al-Taei, Shiroh Futaki, Toshihide Takeuchi, Ikuhiko Nakase, Gert Storm, Arwyn T. Jones
Biochemical Journal Apr 2007, 403 (2) 335-342; DOI: 10.1042/BJ20061808
del.icio.us logo Digg logo Reddit logo Technorati logo Twitter logo CiteULike logo Facebook logo Mendeley logo
Citation Tools
Temperature-, concentration- and cholesterol-dependent translocation of L- and D-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells
Marjan M. Fretz, Neal A. Penning, Saly Al-Taei, Shiroh Futaki, Toshihide Takeuchi, Ikuhiko Nakase, Gert Storm, Arwyn T. Jones
Biochemical Journal Apr 2007, 403 (2) 335-342; DOI: 10.1042/BJ20061808

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print
Alerts

Please log in to add an alert for this article.

Request Permissions
Save to my folders

View Full PDF

 Open in Utopia Docs
  • Tweet Widget
  • Facebook Like

Jump To

  • Article
    • Abstract
    • INTRODUCTION
    • EXPERIMENTAL
    • RESULTS AND DISCUSSION
    • Acknowledgments
    • References
  • Figures
  • Info & Metrics
  • Supplementary Data
  • PDF

Keywords

cell-penetrating peptide
endocytosis
fluorescence microscopy
KG1a cell line
leukaemia
octa-arginine

Related Articles

Cited By...

Subjects

  • Cell
  • Portland Press Homepage
  • Publish With Us
  • Advertising
  • Technical Support
  • Biochemical Journal
  • Clinical Science
  • Essays in Biochemistry
  • Emerging Topics in Life Sciences
  • Biochemical Society Transactions
  • Neuronal Signaling
  • Bioscience Reports
  • Cell Signalling Biology
  • Biochemical Society Symposia

Portland Press Limited
Charles Darwin House
12 Roger Street
London WC1N 2JU
Tel: +44(0) 20 7685 2410
Fax: +44(0) 20 7685 2469
Email: editorial@portlandpress.com

The Biochemical Society