The PC motif: a novel and evolutionarily conserved sequence involved in interaction between p40phox and p67phox, SH3 domain-containing cytosolic factors of the phagocyte NADPH oxidase
| Autor: | Hideki Sumimoto, Yoshiyuki Sakaki, Tetsuro Ago, Kazuhito Mizuki, Kenichiro Hata, Koichiro Takeshige, Takashi Ito, Shigetaka Kitajima, Rika Nakamura |
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| Rok vydání: | 1998 |
| Předmět: |
Recombinant Fusion Proteins
Molecular Sequence Data Sequence alignment Biology Transfection Biochemistry SH3 domain Conserved sequence Evolution Molecular src Homology Domains Cytosol Humans Point Mutation Amino Acid Sequence Binding site Peptide sequence Conserved Sequence chemistry.chemical_classification Phagocytes NADPH oxidase Binding Sites Sequence Homology Amino Acid NADPH Oxidases Phosphoproteins Amino acid chemistry Amino Acid Substitution biology.protein Mutagenesis Site-Directed Signal transduction Sequence Alignment |
| Zdroj: | European journal of biochemistry. 251(3) |
| ISSN: | 0014-2956 |
| Popis: | The superoxide-generating NADPH oxidase, dormant in resting phagocytes, is activated during phagocytosis following assembly of the membrane-integrated protein cytochrome b558 and cytosolic factors. Among the latter are the three proteins containing Src homology 3 (SH3) domains, p67phox, p47phox and p40phox. While the first two factors are indispensable for the activity, p40phox is tightly associated with p67phox in resting cells and is suggested to have some modulatory role. Here we describe a systematic analysis of the interaction between p40phox and p67phox using the yeast two-hybrid system and in vitro binding assays with recombinant proteins. Both methods unequivocally showed that the minimum requirements for stable interaction are the C-terminal region of p40phox and the region between the two SH3 domains of p67phox. This interaction is maintained even in the presence of anionic amphiphiles used for the activation of the NADPH oxidase, raising a possibility that it mediates constitutive association of the two factors in both resting and activated cells. The C-terminal region of p40phox responsible for the interaction contains a characteristic stretch of amino acids designated as the PC motif, that also exists in other signal-transducing proteins from yeast to human. Intensive site-directed mutagenesis to the motif in p40phox revealed that it plays a critical role in the binding to p67phox. Thus the PC motif appears to represent a novel module for protein-protein interaction used in a variety of signaling pathways. |
| Databáze: | OpenAIRE |
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