Synthesis and localization of myeloperoxidase protein in transfected BHK cells.
Processing and localization of myeloperoxidase was studied in nonmyeloid cells. For this purpose BHK cells were transfected with human myeloperoxidase cDNA. In the transfected cells a protein with mol wt of 85,000 was found, which reacted with the specific anti-human myeloperoxidase antiserum. In size and in sensitivity to endo-beta-N-acetylglucosaminidase H this protein resembled the myeloperoxidase precursor synthesized in human promyelocytes. Unlike in the promyelocytes, in BHK cells the 85,000-Da protein was not converted to 60,000- and 14,000-Da polypeptides of the mature enzyme. In Percoll gradients the protein was found predominantly in the light membrane fractions. Microscopic examination revealed a conspicuous immune reaction over the endoplasmic reticulum and nuclear membranes and a moderate labeling over lysosome-like organelles. Pulse-chase experiments indicated that the protein was slowly released from the endoplasmic reticulum; after 1 day the protein was found in similar amounts in cells and in the medium. The secreted protein contained at least one endo-beta-N-acetylglucosaminidase-resistant oligosaccharide. It is suggested that normal intracellular segregation of myeloperoxidase depends on a signal or component, which is not or incompletely expressed in BHK cells.
Cully
J
J
Institut für Physiologische Chemie und Pathobiochemie, Westfälische Wilhelms-Universität Münster, Federal Republic of Germany.
Harrach
B
B
Hauser
H
H
Harth
N
N
Robenek
H
H
Nagata
S
S
Hasilik
A
A
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D006801:Humans; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure; D015473:Leukemia, Promyelocytic, Acute / Q000201:enzymology; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D009195:Peroxidase / Q000096:biosynthesis* / Q000302:isolation & purification; D013347:Subcellular Fractions / Q000201:enzymology / Q000648:ultrastructure; D014162:Transfection
D010455:Peptides; D009195:Peroxidase