Coordinated participation of calreticulin and calnexin in the biosynthesis of myeloperoxidase.
Myeloperoxidase (MPO) is a neutrophil lysosomal hemeprotein essential for optimal oxygen-dependent microbicidal activity. We have demonstrated previously that calreticulin, a luminal endoplasmic reticulum protein, functions as a molecular chaperone during myeloperoxidase biosynthesis, associating reversibly with the heme-free precursor apopro-MPO. Because the membrane-bound endoplasmic reticulum protein calnexin is structurally and functionally related to calreticulin, we assessed the role of calnexin in myeloperoxidase biosynthesis. Like calreticulin, calnexin coprecipitated exclusively with glycosylated MPO precursors and with apopro-MPO but, in contrast to calreticulin, also with the enzymatically active, heme-containing precursor pro-MPO. To determine if calnexin participated in heme insertion into MPO, we compared the kinetics of chaperone association with MPO precursors using stable transfectants expressing cDNA encoding wild type MPO or mutated forms that do not acquire heme. Transfectants expressing mutant cDNA had prolonged association of MPO-related precursors with calreticulin and especially with calnexin. These studies demonstrate that 1) both calreticulin and calnexin associated with glycosylated apopro-MPO; 2) only calnexin associated selectively with the enzymatically active, heme-containing precursor pro-MPO; and 3) mutants unable to incorporate heme had prolonged association with calnexin. These findings represent the first evidence of a specialized role for calnexin in facilitating protein maturation in the endoplasmic reticulum of myeloid cells.
Nauseef
W M
WM
Inflammation Program and Departments of Medicine, University of Iowa and Veterans Affairs Medical Center, Iowa City, Iowa 52242, USA. william-nauseef@uiowa.edu
McCormick
S J
SJ
Goedken
M
M
D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D037282:Calreticulin; D004792:Enzyme Precursors / Q000378:metabolism; D006801:Humans; D009195:Peroxidase / Q000096:biosynthesis*; D012261:Ribonucleoproteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D004792:Enzyme Precursors; D012261:Ribonucleoproteins; D037281:Calnexin; D009195:Peroxidase