Tyrosine sulfation in N-terminal domain of human C5a receptor is necessary for binding of chemotaxis inhibitory protein of Staphylococcus aureus
| Autor: | Yingchun Xu, Zhen-jia Liu, Lei Jiang, Ai-xia Wang, Jinming Gao, Guanhua Du, Yan-juan Yang |
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| Rok vydání: | 2011 |
| Předmět: |
Tyrosine sulfation
Staphylococcus aureus chemical and pharmacologic phenomena Plasma protein binding Biology Inhibitory postsynaptic potential medicine.disease_cause C5a receptor Bacterial Proteins medicine Humans Pharmacology (medical) Tyrosine Receptor Anaphylatoxin C5a Cell Line Transformed Pharmacology HEK 293 cells hemic and immune systems Chemotaxis General Medicine Protein Structure Tertiary Receptors Complement HEK293 Cells Biochemistry Original Article Protein Binding |
| Zdroj: | Acta pharmacologica Sinica. 32(8) |
| ISSN: | 1745-7254 |
| Popis: | Staphylococcus aureus evades host defense through releasing several virulence proteins, such as chemotaxis inhibitory protein of staphylococcus aureus (CHIPS). It has been shown that extracellular N terminus of C5a receptor (C5aR) forms the binding domain for CHIPS, and tyrosine sulfation is emerging as a key factor in determining protein-protein interaction. The aim of this study was to evaluate the role of tyrosine sulfation of N-terminal of C5aR in its binding with CHIPS.Expression plasmids encoding C5aR and its mutants were prepared using PCR and site-directed mutagenesis and were used to transfect HEK 293T cells using calcium phosphate. Recombinant CHIPS protein was purified. Western blotting was used to examine the binding efficiency of CHIPS to C5aR or its mutants.CHIPS exclusively binds to C5aR, but not to C5L2 or C3aR. A nonspecific sulfation inhibitor, sodium chlorate (50 nmol/L), diminishes the binding ability of C5aR with CHIPS. Blocking sulfation by mutation of tyrosine to phenylalanine at positions 11 and 14 of C5aR N terminus, which blocked sulfation, completely abrogates CHIPS binding. When tyrosine 14 alone was mutated to phenylalanine, the binding efficiency of recombinant CHIPS was substantially decreased.The results demonstrate a structural basis of C5aR-CHIPS association, in which tyrosine sulfation of N-terminal C5aR plays an important role. Our data may have potential significance in development of novel drugs for therapeutic intervention. |
| Databáze: | OpenAIRE |
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