Localization of serglycin in human neutrophil granulocytes and their precursors.
Serglycin is a major proteoglycan of hematopoietic cells. It is thought to play a role in the packaging of granule proteins in human neutrophil granulocytes. The presence of serglycin in myeloid cells has been demonstrated only at the transcriptional level. We generated a polyclonal antibody against recombinant human serglycin. Here, we show the localization of serglycin in humans during neutrophil differentiation. Immunocytochemistry revealed serglycin immunoreactivity in the Golgi area of promyelocytes (PM) and myelocytes (MC), as well as in a few band cells and mature neutrophil granulocytes. Granular staining was detected near the Golgi apparatus in some of the PM, and the major part of the cytoplasm was negative. Immunoelectron microscopy showed serglycin immunoreactivity located to the Golgi apparatus and a few immature granules of PM and MC. The decreasing level of serglycin protein during myeloid differentiation coincided with a decrease of mRNA expression, as evaluated by Northern blotting. Subcellular fractions of neutrophil granulocytes were obtained. Serglycin immunoreactivity was detected in the fraction containing Golgi apparatus, plasma membrane, and secretory vesicles by Western blotting and enzyme-linked immunosorbent assay. Serglycin was not detected in subcellular fractions containing primary, secondary, or tertiary granules. Together, these findings indicate that serglycin is located to the Golgi apparatus and a few immature granules during neutrophil differentiation. This is consistent with a function for serglycin in formation of granules in neutrophil granulocytes. Our findings contrast the view that native serglycin is present in mature granules and plays a role in packaging and regulating the activity of proteolytic enzymes there.
Niemann
Carsten Utoft
CU
Rigshospitalet, Department of Haematology, Granulocytlaboratoriet, Building 9322, Blegdamsvej 9, DK-2100, Copenhagen, Denmark. niemann@dadlnet.dk
Cowland
Jack Bernard
JB
Klausen
Pia
P
Askaa
Jon
J
Calafat
Jero
J
Borregaard
Niels
N
D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D011509:Proteoglycans / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D033921:Vesicular Transport Proteins
D011509:Proteoglycans; D012333:RNA, Messenger; D033921:Vesicular Transport Proteins; C075430:serglycin