Cloning and sequencing of rabbit leukocyte NADPH oxidase genes reveals a unique p67(phox) homolog.
The NADPH oxidase plays an important role in immune and nonimmune cell functions. Because rabbits represent an established model for studying a number of important disease processes that involve NADPH oxidase activity, we carried out studies to clone and sequence all five rabbit leukocyte NADPH oxidase genes. Comparison of the rabbit sequences with those of other species showed that, with the exception of p67(phox), the rabbit phox proteins were highly conserved. In contrast, rabbit p67(phox) had a very divergent C-terminus and was 17 amino acids longer than any other known p67(phox) homolog. This was surprising, given the high degree of conservation among all of the phox proteins sequenced previously. To evaluate the functional consequences of this difference, wild-type rabbit p67(phox) and a mutated rabbit p67(phox) missing the C-terminal 17 amino acids were expressed and analyzed in a cell-free assay. Our results show that the full-length and truncated rabbit p67(phox) proteins were able to support oxidase activity, although the truncated form reproducibly supported a higher level of activity than full-length p67(phox). These studies contribute to our understanding of the nature of the leukocyte NADPH oxidase in different species and will be valuable in future research using the rabbit model.
Gauss
Katherine A
KA
Department of Veterinary Molecular Biology, Montana State University, Bozeman, 59717, USA.
Mascolo
Patrice L
PL
Siemsen
Daniel W
DW
Nelson
Laura K
LK
Bunger
Peggy L
PL
Pagano
Patrick J
PJ
Quinn
Mark T
MT
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D007962:Leukocytes / Q000201:enzymology*; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics*; D011817:Rabbits; D016415:Sequence Alignment; D013045:Species Specificity
D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D019255:NADPH Oxidases