Involvement of syntaxin 4 in the transport of membrane-type 1 matrix metalloproteinase to the plasma membrane in human gastric epithelial cells.
Membrane-type 1 matrix metalloproteinase (MT1-MMP) localized on the plasma membrane plays a central role in various normal biological responses including tissue remodeling, wound heeling, and angiogenesis and in cancer cell invasion and metastasis, by functioning as a collagenase and activating other matrix metalloproteinases. In order to elucidate the molecular mechanism of the MT1-MMP targeted localization on the plasma membrane, we examined the participation of syntaxin proteins in MT1-MMP intracellular transport to the plasma membrane in human gastric epithelial AGS cells. Western blotting showed that syntaxin 3 and 4 proteins, which are known to function in intracellular transport towards the plasma membrane, were expressed in AGS cells. Immunocytochemistry revealed that transient transfection of AGS cells with dominant-negative mutant syntaxin 4 decreased plasma membrane MT1-MMP expression. In contrast, transient transfection with either dominant-negative mutant syntaxin 3 or 7 did not affect MT1-MMP localization on the plasma membrane. Cell surface biotinylation assay and Matrigel chamber assay demonstrated that stable transfection with dominant-negative mutant syntaxin 4 decreased the amount of MT1-MMP on the plasma membranes and inhibited the cell invasiveness. We suggest that syntaxin 4 is involved in the intracellular transport of MT1-MMP toward the plasma membrane.
Miyata
Tomohiko
T
Department of Gastroenterology, Jichi Medical School, Tochigi 329-0498, Japan.
Ohnishi
Hirohide
H
Suzuki
Junko
J
Yoshikumi
Yukako
Y
Ohno
Hideki
H
Mashima
Hirosato
H
Yasuda
Hiroshi
H
Ishijima
Takako
T
Osawa
Hiroyuki
H
Satoh
Kiichi
K
Sunada
Keijiro
K
Kita
Hiroto
H
Yamamoto
Hironori
H
Sugano
Kentaro
K
D001692:Biological Transport; D019921:Biotinylation; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D003094:Collagen / Q000494:pharmacology; D004247:DNA / Q000737:chemistry; D004338:Drug Combinations; D005799:Genes, Dominant; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000494:pharmacology; D053504:Matrix Metalloproteinases, Membrane-Associated; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008666:Metalloendopeptidases / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D050765:Qa-SNARE Proteins; D014162:Transfection
D004338:Drug Combinations; D007797:Laminin; D008565:Membrane Proteins; D011509:Proteoglycans; D050765:Qa-SNARE Proteins; C059485:matrigel; D003094:Collagen; D004247:DNA; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases