Human SCAMP5, a novel secretory carrier membrane protein, facilitates calcium-triggered cytokine secretion by interaction with SNARE machinery.
Cytokines produced by immune cells play pivotal roles in the regulation of both innate and adaptive immunity. However, the mechanisms controlling secretion of cytokines have not been fully elucidated. Secretory carrier membrane proteins (SCAMPs) are widely distributed integral membrane molecules implicated in regulating vesicular transport. In this study, we report the functional characterization of human SCAMP5 (hSCAMP5), a novel SCAMP protein that is widely expressed by a variety of neuronal and nonneuronal tissues and cells. By measuring the cytokine secretion (RANTES/CCL5 and IL-1beta) as an exocytotic model, we show that hSCAMP5 can promote the calcium-regulated signal peptide-containing cytokine (CCL5 but not IL-1beta) secretion in human epithelial cancer cells, human monocytes, and mouse macrophages. By using subcellular fractionation, immunofluorescence confocal microscopy, and membrane vesicle immunoisolation methods, we find that hSCAMP5 is mainly localized in the Golgi-associated compartments, and the calcium ionophore ionomycin can trigger a rapid translocation of hSCAMP5 from Golgi apparatus to plasma membrane along the classical exocytosis pathway. During the translocation of hSCAMP5 from Golgi apparatus to plasma membrane, hSCAMP5 can codistribute and complex with local soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs) molecules. We further demonstrate that hSCAMP5 can directly interact with the calcium sensor synaptotagmins via the cytosolic C-terminal tail of hSCAMP5, thus providing a potential molecular mechanism linking SCAMPs with the SNARE molecules. Our findings suggest that hSCAMP5, in cooperation with the SNARE machinery, is involved in calcium-regulated exocytosis of signal peptide-containing cytokines.
Han
Chaofeng
C
Institute of Immunology, Tsinghua University School of Medicine, Beijing, People's Republic of China.
Chen
Taoyong
T
Yang
Mingjin
M
Li
Nan
N
Liu
Haibo
H
Cao
Xuetao
X
D002118:Calcium / Q000502:physiology*; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D005089:Exocytosis / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D021382:Protein Sorting Signals / Q000502:physiology; D011956:Receptors, Cell Surface / Q000378:metabolism / Q000502:physiology*; D050600:SNARE Proteins / Q000737:chemistry / Q000378:metabolism*; D022142:Secretory Vesicles / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology
D002352:Carrier Proteins; D016207:Cytokines; D008565:Membrane Proteins; D021382:Protein Sorting Signals; D011956:Receptors, Cell Surface; C540013:SCAMP5 protein, human; D050600:SNARE Proteins; C091829:secretory component receptor, human; D002118:Calcium