The CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells.

2.50
Hdl Handle:
http://hdl.handle.net/10033/124109
Title:
The CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells.
Authors:
Kaur, Simran Jeet; Nerlich, Andreas; Bergmann, Simone; Rohde, Manfred ( 0000-0003-0522-3580 ) ; Fulde, Marcus; Zähner, Dorothea; Hanski, Emanuel; Zinkernagel, Annelies; Nizet, Victor; Chhatwal, Gursharan S; Talay, Susanne R
Abstract:
Streptococcus pyogenes expresses the LPXTG motif-containing cell envelope serine protease SpyCep (also called ScpC, PrtS) that degrades and inactivates the major chemoattractant interleukin 8 (IL-8), thereby impairing host neutrophil recruitment. In this study, we identified a novel function of SpyCep: the ability to mediate uptake into primary human endothelial cells. SpyCep triggered its uptake into endothelial cells but not into human epithelial cells originating from pharynx or lung, indicating an endothelial cell-specific uptake mechanism. SpyCep mediated cellular invasion by an endosomal/lysosomal pathway distinct from the caveolae-mediated invasion pathway of S. pyogenes. Recombinant expression and purification of proteolytically active SpyCep and a series of subfragments allowed functional dissection of the domains responsible for endothelial cell invasion and IL-8 degradation. The N-terminal PR domain was sufficient to mediate endothelial cell invasion, whereas for IL-8-degrading activity, the protease domain and the flanking A domain were required. A polyclonal rabbit serum raised against the recombinant protease efficiently blocked the invasion-mediating activity of SpyCep but not its proteolytic function, further indicating that SpyCep-mediated internalization is independent from its enzymatic activity. SpyCep may thus specifically mediate its own uptake as secreted protein into human endothelial cells.
Affiliation:
Department of Microbial Pathogenesis, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.
Citation:
The CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells. 2010, 285 (36):27798-805 J. Biol. Chem.
Journal:
The Journal of biological chemistry
Issue Date:
3-Sep-2010
URI:
http://hdl.handle.net/10033/124109
DOI:
10.1074/jbc.M109.098053
PubMed ID:
20562101
Type:
Article
Language:
en
ISSN:
1083-351X
Appears in Collections:
Publications of Dept. Medizinische Mikrobiologie (MMIK)

Full metadata record

DC FieldValue Language
dc.contributor.authorKaur, Simran Jeeten
dc.contributor.authorNerlich, Andreasen
dc.contributor.authorBergmann, Simoneen
dc.contributor.authorRohde, Manfreden
dc.contributor.authorFulde, Marcusen
dc.contributor.authorZähner, Dorotheaen
dc.contributor.authorHanski, Emanuelen
dc.contributor.authorZinkernagel, Anneliesen
dc.contributor.authorNizet, Victoren
dc.contributor.authorChhatwal, Gursharan Sen
dc.contributor.authorTalay, Susanne Ren
dc.date.accessioned2011-03-10T12:34:53Zen
dc.date.available2011-03-10T12:34:53Zen
dc.date.issued2010-09-03en
dc.identifier.citationThe CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells. 2010, 285 (36):27798-805 J. Biol. Chem.en
dc.identifier.issn1083-351Xen
dc.identifier.pmid20562101en
dc.identifier.doi10.1074/jbc.M109.098053en
dc.identifier.urihttp://hdl.handle.net/10033/124109en
dc.description.abstractStreptococcus pyogenes expresses the LPXTG motif-containing cell envelope serine protease SpyCep (also called ScpC, PrtS) that degrades and inactivates the major chemoattractant interleukin 8 (IL-8), thereby impairing host neutrophil recruitment. In this study, we identified a novel function of SpyCep: the ability to mediate uptake into primary human endothelial cells. SpyCep triggered its uptake into endothelial cells but not into human epithelial cells originating from pharynx or lung, indicating an endothelial cell-specific uptake mechanism. SpyCep mediated cellular invasion by an endosomal/lysosomal pathway distinct from the caveolae-mediated invasion pathway of S. pyogenes. Recombinant expression and purification of proteolytically active SpyCep and a series of subfragments allowed functional dissection of the domains responsible for endothelial cell invasion and IL-8 degradation. The N-terminal PR domain was sufficient to mediate endothelial cell invasion, whereas for IL-8-degrading activity, the protease domain and the flanking A domain were required. A polyclonal rabbit serum raised against the recombinant protease efficiently blocked the invasion-mediating activity of SpyCep but not its proteolytic function, further indicating that SpyCep-mediated internalization is independent from its enzymatic activity. SpyCep may thus specifically mediate its own uptake as secreted protein into human endothelial cells.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshAntibodiesen
dc.subject.meshCell Lineen
dc.subject.meshCloning, Molecularen
dc.subject.meshEndocytosisen
dc.subject.meshEndosomesen
dc.subject.meshEndothelial Cellsen
dc.subject.meshHumansen
dc.subject.meshInterleukin-8en
dc.subject.meshLysosomesen
dc.subject.meshPeptide Hydrolasesen
dc.subject.meshProtein Structure, Tertiaryen
dc.subject.meshProtein Transporten
dc.subject.meshStreptococcus pyogenesen
dc.titleThe CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells.en
dc.typeArticleen
dc.contributor.departmentDepartment of Microbial Pathogenesis, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.en
dc.identifier.journalThe Journal of biological chemistryen

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