Expression of azurophil and specific granule proteins during differentiation of NB4 cells in neutrophils.
Neutrophils contain several populations of secretory granules with characteristic sets of proteins. Granule proteins are sorted into their respective granule types by temporal regulation of their expression during cell differentiation and/or by specific targeting signals. We investigated the expression of some granule proteins in human promyelocytic NB4 cells. Like other myeloid cell lines which can be differentiated into neutrophils, NB4 cells lack the specific-granule population. We report here that, nevertheless, they express the specific-granule matrix protein lactoferrin, when differentiated with retinoic acid. Lactoferrin and the azurophil-granule protein beta-glucuronidase were simultaneously expressed, whereas myeloperoxidase expression had stopped, showing that azurophil-granule proteins are not all produced concomitantly. Cell fractionation by Percoll gradient revealed that while beta-glucuronidase co-fractionated with myeloperoxidase, lactoferrin was mostly contained in a vesicular compartment free of markers for azurophil granules, plasma membrane, and Golgi. This vesicular compartment was not implicated in regulated exocytosis since it was not mobilized by secretagogues, which, in parallel, induced the release of myeloperoxidase. Furthermore, the specific granule-membrane protein cytochrome b558 also became expressed during NB4-cell differentiation. However, it did not co-localize with lactoferrin but was present in the plasma-membrane fraction. Therefore, differentiation of NB4 cells with retinoic acid leads to the expression of specific- and azurophil-granule proteins and provides a unique cell line model to study the mechanisms involved in the sorting of azurophil- and specific-granule proteins.
Grégoire
C
C
Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique, Toulouse, France.
Welch
H
H
Astarie-Dequeker
C
C
Maridonneau-Parini
I
I
D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000502:physiology*; D002458:Cell Fractionation; D003573:Cytochrome b Group / Q000032:analysis; D003594:Cytoplasmic Granules / Q000201:enzymology / Q000378:metabolism*; D005966:Glucuronidase / Q000032:analysis; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism; D019255:NADPH Oxidases; D009504:Neutrophils / Q000502:physiology; D009195:Peroxidase / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D015415:Biomarkers; D003573:Cytochrome b Group; D014212:Tretinoin; C036888:cytochrome b558; D009195:Peroxidase; D019255:NADPH Oxidases; D005966:Glucuronidase; D007781:Lactoferrin