Calreticulin functions as a molecular chaperone in the biosynthesis of myeloperoxidase.
Myeloperoxidase (MPO), a lysosomal heme protein found exclusively in neutrophils and monocytes, is necessary for efficient oxygen-dependent microbicidal activity. Acquisition of heme by the heme-free MPO precursor apopro-MPO appears to be a prerequisite for its subsequent proteolytic processing and advancement along the biosynthetic pathway to mature MPO. We present data indicating that calreticulin (CRT), a high capacity calcium-binding protein residing in the lumen of the endoplasmic reticulum of a wide variety of cells, interacts specifically with fully glycosylated apopro-MPO. Biosynthetically radiolabeled CRT (60 kDa) and apopro-MPO (90 kDa) were coprecipitated from PLB 985 cells by monospecific antiserum against CRT when the immunoprecipitations were performed either under nondenaturing conditions or following reversible crosslinking. Nonglycosylated MPO precursors synthesized in the presence of tunicamycin did not interact with CRT. The CRT-apopro-MPO interaction was restricted to an early phase of MPO biosynthesis, and CRT did not interact with the later appearing, heme-containing species of MPO, i.e. pro-MPO or the heavy subunit of mature MPO. These data show that CRT participates in the post-translational processing of MPO, perhaps by maintaining apopro-MPO in a conformation competent to accommodate insertion of the heme group. In this general way, CRT shares certain functional properties with the structurally homologous transmembrane calcium-binding endoplasmic reticulum protein calnexin. Both interact with glycosylated biosynthetic precursors of proteins selectively expressed in specialized cells.
Nauseef
W M
WM
Department of Medicine, Veterans Administration Medical Center, Iowa City, Iowa.
McCormick
S J
SJ
Clark
R A
RA
D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D018833:Chaperonins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004792:Enzyme Precursors / Q000378:metabolism; D006801:Humans; D009195:Peroxidase / Q000096:biosynthesis*; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D004792:Enzyme Precursors; D011506:Proteins; D012261:Ribonucleoproteins; D009195:Peroxidase; D018833:Chaperonins