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 p41^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.