Trafficking of human ADAM 12-L: retention in the trans-Golgi network.
We have investigated the trafficking of the membrane-anchored form of human ADAM 12 (ADAM 12-L) fused to a green fluorescence protein tag. Subcellular localization of the protein in transiently transfected cells was determined by fluorescence microscopy and trypsin sensitivity. Full-length ADAM 12-L was retained in a perinuclear compartment, which was shown to be the trans-Golgi network. In contrast, ADAM 12-L lacking the cytoplasmic domain reached the cell surface. Based on analysis of deletions and mutations of the cytoplasmic tail of ADAM 12-L, the retention signal is comprised of both the cytoplasmic and transmembrane domains, but not the Src homology 3 domain (SH3) binding sites. These results raise the possibility that a trafficking checkpoint in the trans-Golgi network is one of the cellular mechanisms for regulation of ADAM 12-L function, by allowing a rapid release of ADAM 12-L to the cell surface under specific stimuli.
Hougaard
S
S
Institute of Molecular Pathology, University of Copenhagen, Frederik V's vej 11, Copenhagen, DK-2100, Denmark.
Loechel
F
F
Xu
X
X
Tajima
R
R
Albrechtsen
R
R
Wewer
U M
UM
D051722:ADAM Proteins; D000072199:ADAM12 Protein; D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D019556:COS Cells; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003593:Cytoplasm / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000096:biosynthesis / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D017830:Octoxynol
D008565:Membrane Proteins; D017830:Octoxynol; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C000606480:ADAM12 protein, human; D008666:Metalloendopeptidases