Cloning Of Rat P47(Phox) And Comparison With Human P47(Phox)

A cDNA encoding rat p47(phox) was cloned from rat spleen cDNA library, utilizing rapid amplification of cDNA ends. The open reading frame corresponded to 389 amino acids: It contained the phagocyte oxidase homology domain, two Src homology 3 domains and a proline rich region, all of which are conserved in mammalian p47(phox) sequences. Rat p47(phox) displayed the highest degree of identity to mouse p47(phox) (94%). We expressed and purified rat p47(phox) as a glutathione S-transferase fusion protein, and found that the rat protein could replace human p47(phox) in a cell-free activation system for human NADPH oxidase, giving about half activity. Although rat 12-lipoxygenase interacted with human p47(phox) in a yeast two-hybrid system, this was not the case for rat p47(phox).