A novel cell penetrating aspartic protease inhibitor blocks processing and presentation of tetanus toxoid more efficiently than pepstatin A

Nousheen Zaidi, Timo Burster, Vinod Sommandas, Timo Herrmann, Bernhard O Boehm, Christoph Driessen, Wolfgang Voelter, Hubert Kalbacher

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Selective inhibition of enzymes involved in antigen processing such as cathepsin E and cathepsin D is a valuable tool for investigating the roles of these enzymes in the processing pathway. However, the aspartic protease inhibitors, including the highly potent pepstatin A (PepA), are inefficiently transported across the cell membrane and thus have limited access to antigen processing compartments. Previously described mannose-pepstatin conjugates were efficiently taken up by the cells via receptor mediated uptake. However, cells without mannose receptors are unable to take up these conjugates efficiently. The aim of the present study was to synthesize new cell-permeable aspartic protease inhibitors by conjugating pepstatin A with well-known cell penetrating peptides (CPPs). To achieve this, the most commonly used CPPs namely pAntp(43-58) (penetratin), Tat(49-60), and 9-mer of l-arginine (R9), were synthesized and coupled to pepstatin. The enzyme inhibitory properties of these bioconjugates and their cellular uptake into MCF7 (human breast cancer cell line), Boleths (EBV-transformed B-cell line) and dendritic cells (DC) were the focus of our study. We found that the bioconjugate PepA-penetratin (PepA-P) was the most efficient cell-permeable aspartic protease inhibitor tested, and was more efficient than unconjugated PepA. Additionally, we found that PepA-P efficiently inhibited the tetanus toxoid C-fragment processing in peripheral blood mononuclear cells (PBMC), primary DC and in primary B cells. Therefore, PepA-P can be used in studying the role of intracellular aspartic proteases in the MHC class II antigen processing pathway. Moreover, inhibition of tetanus toxoid C-fragment processing by PepA-P clearly implicates the role of aspartic proteinases in antigen processing.

Original languageEnglish
Pages (from-to)243-9
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume364
Issue number2
DOIs
Publication statusPublished - Dec 14 2007

Keywords

  • Antigen Presentation
  • Arginine
  • Aspartic Acid Endopeptidases
  • B-Lymphocytes
  • Carrier Proteins
  • Cell Membrane Permeability
  • Cells, Cultured
  • Dendritic Cells
  • Gene Products, tat
  • Humans
  • Pepstatins
  • Peptide Fragments
  • Structure-Activity Relationship
  • Tetanus Toxoid
  • Journal Article
  • Research Support, Non-U.S. Gov't

Fingerprint Dive into the research topics of 'A novel cell penetrating aspartic protease inhibitor blocks processing and presentation of tetanus toxoid more efficiently than pepstatin A'. Together they form a unique fingerprint.

  • Cite this