Document Detail


A novel cell penetrating aspartic protease inhibitor blocks processing and presentation of tetanus toxoid more efficiently than pepstatin A.
MedLine Citation:
PMID:  17937927     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Nousheen Zaidi; Timo Burster; Vinod Sommandas; Timo Herrmann; Bernhard O Boehm; Christoph Driessen; Wolfgang Voelter; Hubert Kalbacher
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-10-04
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  364     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2007 Dec 
Date Detail:
Created Date:  2007-10-31     Completed Date:  2008-01-25     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  243-9     Citation Subset:  IM    
Affiliation:
Medical and Natural Sciences Research Centre, University of Tübingen, Ob dem Himmelreich 7, 72074 Tübingen, Germany.
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MeSH Terms
Descriptor/Qualifier:
Antigen Presentation / drug effects
Arginine / chemistry,  pharmacology*
Aspartic Acid Endopeptidases / antagonists & inhibitors*,  metabolism
B-Lymphocytes / drug effects,  immunology
Carrier Proteins / chemistry,  pharmacology*
Cell Membrane Permeability
Cells, Cultured
Dendritic Cells / drug effects,  immunology
Gene Products, tat / chemistry*
Humans
Pepstatins / chemistry,  pharmacology*
Peptide Fragments / chemistry,  pharmacology*
Structure-Activity Relationship
Tetanus Toxoid / immunology*
Chemical
Reg. No./Substance:
0/Carrier Proteins; 0/Gene Products, tat; 0/Pepstatins; 0/Peptide Fragments; 0/Tetanus Toxoid; 0/penetratin; 39324-30-6/pepstatin; 74-79-3/Arginine; EC 3.4.23.-/Aspartic Acid Endopeptidases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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