Endocytosis targets exogenous material selectively to cathepsin S in live human dendritic cells, while cell-penetrating peptides mediate nonselective transport to cysteine cathepsins

Reich, Michael; Swieten, Paul F. van; Sommandas, Vinod; Kraus, Marianne; Fischer, Rainer; Weber, Ekkehard; Kalbacher, Hubert; Overkleeft, Herman S.

doi:10.1189/jlb.1006600

Cited by 24 publications

(16 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, these pathways might not be as general and could depend on the specific sequence of the CPP and the cargo attached to it. Several recent reports indicate that these peptides get inserted into the cell through endocytosis and pinocytosis [60][61][62][63][64]. We observed in our simulations that these peptides interact very strongly with the membrane, introducing lipid rearrangements and stress.…”

Section: Discussionsupporting

confidence: 52%

Cell Penetrating Peptides: How Do They Do It?

2007

View full text Add to dashboard Cite

Cell penetrating peptides consist of short sequences of amino acids containing a large net positive charge that are able to penetrate almost any cell, carrying with them relatively large cargoes such as proteins, oligonucleotides, and drugs. During the 10 years since their discovery, the question of how they manage to translocate across the membrane has remained unanswered. The main discussion has been centered on whether they follow an energy-independent or an energy-dependent pathway. Recently, we have discovered the possibility of an energy-independent pathway that challenges fundamental concepts associated with protein-membrane interactions (Herce and Garcia, PNAS, 104: 20805 (2007) [1]). It involves the translocation of charged residues across the hydrophobic core of the membrane and the passive diffusion of these highly charged peptides across the membrane through the formation of aqueous toroidal pores. The aim of this review is to discuss the details of the mechanism and interpret some experimental results consistent with this view.

show abstract

Section: Discussionsupporting

confidence: 52%

Cell Penetrating Peptides: How Do They Do It?

2007

View full text Add to dashboard Cite

show abstract

“…DCG0N, a derivative of DCG04 with identical labeling characteristics, is a biotin-labeled active site-directed probe specific for active papain-like proteases (27,28). Subcellular fractions, in which protease activity was inhibited, were preincubated for 1 h at 37°C with either E64 at a concentration of 10 M or LHVS at a concentration of 10 nM.…”

Section: Methodsmentioning

confidence: 99%

Quantifying Cathepsin S Activity in Antigen Presenting Cells Using a Novel Specific Substrate

Lützner

Kalbacher

2008

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Cathepsin S (CatS) is a lysosomal cysteine protease belonging to the papain superfamily. Because of the relatively broad substrate specificity of this family, a specific substrate for CatS is not yet known. Based on a detailed study of the CatS endopeptidase specificity, using six series of internally quenched fluorescent peptides, we were able to design a specific substrate for CatS. The peptide series was based on the sequence GRWHTVGL-RWE-Lys(Dnp)-DArg-NH 2 , which shows only one single cleavage site between Gly and Leu and where every substrate position between P-3 and P-3 was substituted with up to 15 different amino acids. The endopeptidase specificity of CatS was mainly determined by the P-2, P-1, and the P-3 substrate positions. Based on this result, systematically modified substrates were synthesized. Two of these modified substrates, Mca-GRWPP-MGLPWE-Lys(Dnp)-DArg-NH 2 and Mca-GRWHPMGAPWELys(Dnp)-DArg-NH 2 , did not react with the purified cysteine proteases cathepsin B (CatB) and cathepsin L (CatL). Using a specific CatS inhibitor, we could further show that these two peptides were not cleaved by endosomal fractions of antigen presenting cells (APCs), when CatS was inhibited and related cysteine proteases cathepsin B, H, L and X were still active. Although aspartic proteases like cathepsin E and cathepsin D were also present, our substrates were suitable to quantify cathepsin S activity specifically in APCs, including B cells, macrophages, and dendritic cells without the use of any protease inhibitor. We find that CatS activity differs significantly not only between the three types of professional APCs but also between endosomal and lysosomal compartments.Lysosomal cysteine proteases of the papain family were long believed to be exclusively involved in nonspecific proteolysis within the lysosome. However, the use of specific protease inhibitors and the study of gene knock-out mice suggested that some of them are involved in many other processes, such as cellular homeostasis, autophagy, apoptosis, and antigen presentation (1-3). Antigen presentation by MHC 2 class II molecules requires the entry of antigens into the endosomal-lysosomal compartment. These antigens are then processed by proteolytic enzymes, of which the lysosomal cysteine proteases of the papain family constitute an important subset. The generated peptides bind to MHC class II molecules, which are then displayed at the surface of professional APCs including macrophages, dendritic cells (DCs), and B cells. The variety of MHC class II peptide products that can activate CD4 ϩ T cells is on the one hand determined by allelic variation in MHC molecule binding specificity and on the other by the identity and level of activity of processing proteases present in APCs. Although CatS is one of the major proteases involved in antigen processing (4 -6), specific determination of its activity in antigen presenting cells is currently complicated because a specific substrate is not yet known. One reason for this is that CatB, CatL, and CatS show relati...

show abstract

“…By contrast, the total amount of GAPDH remained unaffected by HMCV infection. Monocytes were included as a control for the distribution of cathepsins (Reich et al, 2007).…”

Section: Hcmv Infection Reduces the Expression Of Endocytic Proteasesmentioning

confidence: 99%

Human cytomegalovirus infection interferes with major histocompatibility complex type II maturation and endocytic proteases in dendritic cells at multiple levels

Keßler

Reich

Jahn

et al. 2008

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

Human cytomegalovirus (HCMV) infection suppresses cellular immunity and results in viral persistence. Dendritic cells (DCs) are susceptible to HCMV, and the development and immune function of HCMV-infected DCs are impaired in vitro. HCMV-derived proteins interfere with different aspects of major histocompatibility complex type II (MHC II) maturation and function in genetically engineered cellular models. This study directly analysed the effect of HCMV on the MHC II-associated antigen processing and presentation machinery in HCMV-infected human DCs in vitro. HCMV-infected DCs failed to mature newly synthesized MHC II to the final stage of SDS-stable MHC II ab dimer/peptide complexes, in contrast to mock-infected controls. MHC II biosynthesis was delayed and reduced, whilst MHC II stability remained unchanged. MHC II surface expression was decreased in the late phase of HCMV infection. In addition, infected DCs decreased the transcription rate of the MHC II-associated proteases cathepsins S, Z, B, H and L and asparagine-specific endopeptidase (AEP). This translated into reduced protein expression of cathepsins H and S, as well as AEP, and less-efficient proteolytic degradation of a peptide substrate by endocytic proteases from HCMV-infected DCs in vitro. Thus, HCMV infection interferes with MHC II biosynthesis and maturation, as well as with the expression and function of endocytic proteases in infected DCs.

show abstract

Endocytosis targets exogenous material selectively to cathepsin S in live human dendritic cells, while cell-penetrating peptides mediate nonselective transport to cysteine cathepsins

Cited by 24 publications

References 41 publications

Cell Penetrating Peptides: How Do They Do It?

Cell Penetrating Peptides: How Do They Do It?

Quantifying Cathepsin S Activity in Antigen Presenting Cells Using a Novel Specific Substrate

Human cytomegalovirus infection interferes with major histocompatibility complex type II maturation and endocytic proteases in dendritic cells at multiple levels

Contact Info

Product

Resources

About