Premium
Cloning and sequencing of rabbit leukocyte NADPH oxidase genes reveals a unique p67 phox homolog
Author(s) -
Gauss Katherine A.,
Mascolo Patrice L.,
Siemsen Daniel W.,
Nelson Laura K.,
Bunger Peggy L.,
Pagano Patrick J.,
Quinn Mark T.
Publication year - 2002
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1189/jlb.71.2.319
Subject(s) - biology , nadph oxidase , oxidase test , cloning (programming) , gene , amino acid , microbiology and biotechnology , conserved sequence , rabbit (cipher) , clone (java method) , peptide sequence , biochemistry , enzyme , statistics , mathematics , computer science , programming language
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.