Wide distribution of granulophysin epitopes in granules of human tissues.
BACKGROUND
The identification and characterization of granule membrane proteins are becoming increasingly important in understanding the packaging and secretory function of granules and characterizing diseases involving granules. A granule membrane protein, granulophysin, has recently been identified in the membranes of platelet dense granules, organelles that contain stored ADP, ATP, serotonin, and calcium. Antibodies that recognize granulophysin also stain granules of monocytes, neutrophils, and lymphokine activated killer cells.


METHODS
In the present study, the distribution of epitopes recognized by antigranulophysin monoclonal antibodies in human tissues was investigated using immunohistochemistry on paraffin sections. Quantitation of the protein was also performed by enzyme-linked immunosorbent assay. The protein was also analyzed in various tissues using Western blotting.


RESULTS
Granulophysin was localized to the granules of skin melanocytes, neurons, endocrine gland cells, exocrine glands (except mucin producing cells), and surface lining cells. Analysis by Western blots revealed a typical staining pattern for granulophysin in lung, adrenal gland, liver, brain, prostate, and pituitary. Atypical bands were present in the pancreas head (47 kDa) and skeletal muscle (34 kDa). A clear distinction was demonstrated between granulophysin and synaptophysin through both immunochemistry and blotting, despite the known cross-reactivity of these two proteins.


CONCLUSIONS
The findings demonstrate that granulophysin is a widely distributed protein that is frequently associated with granules. We speculate that it may be critical in granule function.
Hatskelzon
L
L
Manitoba Institute of Cell Biology, Winnipeg, Canada.
Dalal
B I
BI
Shalev
A
A
Robertson
C
C
Gerrard
J M
JM
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D015153:Blotting, Western; D003594:Cytoplasmic Granules / Q000737:chemistry* / Q000648:ultrastructure; D004702:Endocrine Glands / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000737:chemistry; D007985:Leydig Cells / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D008221:Lymphoid Tissue / Q000737:chemistry; D008297:Male; D008544:Melanocytes / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology; D009132:Muscles / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D009474:Neurons / Q000737:chemistry*; D010179:Pancreas / Q000737:chemistry; D010980:Platelet Membrane Glycoproteins / Q000032:analysis / Q000276:immunology*; D016708:Synaptophysin / Q000032:analysis / Q000276:immunology; D060149:Tetraspanin 30
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C554586:CD63 protein, human; D000939:Epitopes; D008565:Membrane Proteins; D010980:Platelet Membrane Glycoproteins; D016708:Synaptophysin; D060149:Tetraspanin 30