Human B lymphoblastoid cells contain distinct patterns of cathepsin activity in endocytic compartments and regulate MHC class II transport in a cathepsin S-independent manner

Alfred Lautwein, Marianne Kraus, Michael Reich, Timo Burster, J Brandenburg, Herman S Overkleeft, Gerold Schwarz, Winfried Kammer, Ekkehard Weber, Hubert Kalbacher, Alfred Nordheim, Christoph Driessen

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Abstract

Endocytic proteolysis represents a major functional component of the major histocompatibility complex class II antigen-presentation machinery. Although transport and assembly of class II molecules in the endocytic compartment are well characterized, we lack information about the pattern of endocytic protease activity along this pathway. Here, we used chemical tools that visualize endocytic proteases in an activity-dependent manner in combination with subcellular fractionation to dissect the subcellular distribution of the major cathepsins (Cat) CatS, CatB, CatH, CatD, CatC, and CatZ as well as the asparagine-specific endoprotease (AEP) in human B-lymphoblastoid cells (BLC). Endocytic proteases were distributed in two distinct patterns: CatB and CatZ were most prominent in early and late endosomes but absent from lysosomes, and CatH, CatS, CatD, CatC, and AEP distributed between late endosomes and lysosomes, suggesting that CatB and CatZ might be involved in the initial proteolytic attack on a given antigen. The entire spectrum of protease activity colocalized with human leukocyte antigen-DM and the C-terminal and N-terminal processing of invariant chain (Ii) in late endosomes. CatS was active in all endocytic compartments. Surprisingly and in contrast with results from dendritic cells, inhibition of CatS activity by leucine-homophenylalanine-vinylsulfone-phenol prevented N-terminal processing of Ii but did not alter the subcellular trafficking or surface delivery of class II complexes, as deferred from pulse-chase analysis in combination with subcellular fractionation and biotinylation of cell-surface protein. Thus, BLC contain distinct activity patterns of proteases in endocytic compartments and regulate the intracellular transport and surface-delivery of class II in a CatS-independent manner.

Original languageEnglish
Pages (from-to)844-55
Number of pages12
JournalJournal of Leukocyte Biology
Volume75
Issue number5
DOIs
Publication statusPublished - May 2004

Fingerprint

cathepsin S
Cathepsins
Cats
Peptide Hydrolases
Endosomes
Asparagine
Lysosomes
Biotinylation
Histocompatibility Antigens Class II
Antigen Presentation
HLA Antigens
Phenol
Major Histocompatibility Complex
Leucine
Dendritic Cells
Proteolysis
Membrane Proteins
Antigens

Keywords

  • Antigen Presentation
  • Antigens, Differentiation, B-Lymphocyte
  • B-Lymphocytes
  • Biological Transport
  • Cathepsins
  • Cell Fractionation
  • Endocytosis
  • Endosomes
  • Histocompatibility Antigens Class II
  • Humans
  • Lysosomes
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Human B lymphoblastoid cells contain distinct patterns of cathepsin activity in endocytic compartments and regulate MHC class II transport in a cathepsin S-independent manner. / Lautwein, Alfred; Kraus, Marianne; Reich, Michael; Burster, Timo; Brandenburg, J; Overkleeft, Herman S; Schwarz, Gerold; Kammer, Winfried; Weber, Ekkehard; Kalbacher, Hubert; Nordheim, Alfred; Driessen, Christoph.

In: Journal of Leukocyte Biology, Vol. 75, No. 5, 05.2004, p. 844-55.

Research output: Contribution to journalArticle

Lautwein, A, Kraus, M, Reich, M, Burster, T, Brandenburg, J, Overkleeft, HS, Schwarz, G, Kammer, W, Weber, E, Kalbacher, H, Nordheim, A & Driessen, C 2004, 'Human B lymphoblastoid cells contain distinct patterns of cathepsin activity in endocytic compartments and regulate MHC class II transport in a cathepsin S-independent manner', Journal of Leukocyte Biology, vol. 75, no. 5, pp. 844-55. https://doi.org/10.1189/jlb.0803367
Lautwein, Alfred ; Kraus, Marianne ; Reich, Michael ; Burster, Timo ; Brandenburg, J ; Overkleeft, Herman S ; Schwarz, Gerold ; Kammer, Winfried ; Weber, Ekkehard ; Kalbacher, Hubert ; Nordheim, Alfred ; Driessen, Christoph. / Human B lymphoblastoid cells contain distinct patterns of cathepsin activity in endocytic compartments and regulate MHC class II transport in a cathepsin S-independent manner. In: Journal of Leukocyte Biology. 2004 ; Vol. 75, No. 5. pp. 844-55.
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AU - Burster, Timo

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AU - Overkleeft, Herman S

AU - Schwarz, Gerold

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KW - Endosomes

KW - Histocompatibility Antigens Class II

KW - Humans

KW - Lysosomes

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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JO - Journal of Leukocyte Biology

JF - Journal of Leukocyte Biology

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