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Enhancement of endothelial cell migration and in vitro tube formation by TAP20, a novel beta 5 integrin-modulating, PKC theta-dependent protein.

Tang S, Gao Y, Ware JA - J. Cell Biol. (1999)

Bottom Line: A full-length cDNA encoding a novel 20-kD protein, whose expression was PKCtheta-dependent, was identified in endothelial cells, cloned, characterized, and designated as theta-associated protein (TAP) 20.An antiintegrin alphavbeta5 antibody prevented these TAP20 effects.The interaction between TAP20 and beta5 integrin cytoplasmic domain was demonstrated by protein coprecipitation and immunoblotting.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Division, Department of Medicine, Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. tang@aecom.yu.edu

ABSTRACT
Migration, proliferation, and tube formation of endothelial cells are regulated by a protein kinase C isoenzyme PKCtheta. A full-length cDNA encoding a novel 20-kD protein, whose expression was PKCtheta-dependent, was identified in endothelial cells, cloned, characterized, and designated as theta-associated protein (TAP) 20. Overexpression of TAP20 decreased cell adhesion and enhanced migration on vitronectin and tube formation in three-dimensional culture. An antiintegrin alphavbeta5 antibody prevented these TAP20 effects. Overexpression of TAP20 also decreased focal adhesion formation in alphavbeta3-deficient cells. The interaction between TAP20 and beta5 integrin cytoplasmic domain was demonstrated by protein coprecipitation and immunoblotting. Thus, the discovery of TAP20, which interacts with integrin beta5 and modulates cell adhesion, migration, and tube formation, further defines a possible pathway to angiogenesis dependent on PKCtheta.

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Related in: MedlinePlus

Enhancement of HUVEC tube formation on matrix gel by TAP20. Transfected cells were sorted with GFP fluorescence and were seeded on top of the matrix gel with complete M199 medium. The photographs were taken using light microscopy (100× view field) with a video TV camera system, at the timepoints indicated.
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Figure 6: Enhancement of HUVEC tube formation on matrix gel by TAP20. Transfected cells were sorted with GFP fluorescence and were seeded on top of the matrix gel with complete M199 medium. The photographs were taken using light microscopy (100× view field) with a video TV camera system, at the timepoints indicated.

Mentions: Since TAP20 modulated cell adhesion and migration, we asked whether overexpressing TAP20 would alter the ability of cells to form tubes on matrix gel. When the GFP-sorted HUVEC cells were cultured in matrix gel, a three-dimensional matrix, tube formation by TAP20 + GFP transfectants was significantly enhanced (Fig. 6). By 6 h, tube structures were observed in the TAP20 + GFP transfectants, but not in the control GFP cells. The number of tubes formed by TAP20 + GFP transfectants appeared to be dramatically increased at all timepoints compared with that by the control cells.


Enhancement of endothelial cell migration and in vitro tube formation by TAP20, a novel beta 5 integrin-modulating, PKC theta-dependent protein.

Tang S, Gao Y, Ware JA - J. Cell Biol. (1999)

Enhancement of HUVEC tube formation on matrix gel by TAP20. Transfected cells were sorted with GFP fluorescence and were seeded on top of the matrix gel with complete M199 medium. The photographs were taken using light microscopy (100× view field) with a video TV camera system, at the timepoints indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2169340&req=5

Figure 6: Enhancement of HUVEC tube formation on matrix gel by TAP20. Transfected cells were sorted with GFP fluorescence and were seeded on top of the matrix gel with complete M199 medium. The photographs were taken using light microscopy (100× view field) with a video TV camera system, at the timepoints indicated.
Mentions: Since TAP20 modulated cell adhesion and migration, we asked whether overexpressing TAP20 would alter the ability of cells to form tubes on matrix gel. When the GFP-sorted HUVEC cells were cultured in matrix gel, a three-dimensional matrix, tube formation by TAP20 + GFP transfectants was significantly enhanced (Fig. 6). By 6 h, tube structures were observed in the TAP20 + GFP transfectants, but not in the control GFP cells. The number of tubes formed by TAP20 + GFP transfectants appeared to be dramatically increased at all timepoints compared with that by the control cells.

Bottom Line: A full-length cDNA encoding a novel 20-kD protein, whose expression was PKCtheta-dependent, was identified in endothelial cells, cloned, characterized, and designated as theta-associated protein (TAP) 20.An antiintegrin alphavbeta5 antibody prevented these TAP20 effects.The interaction between TAP20 and beta5 integrin cytoplasmic domain was demonstrated by protein coprecipitation and immunoblotting.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Division, Department of Medicine, Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. tang@aecom.yu.edu

ABSTRACT
Migration, proliferation, and tube formation of endothelial cells are regulated by a protein kinase C isoenzyme PKCtheta. A full-length cDNA encoding a novel 20-kD protein, whose expression was PKCtheta-dependent, was identified in endothelial cells, cloned, characterized, and designated as theta-associated protein (TAP) 20. Overexpression of TAP20 decreased cell adhesion and enhanced migration on vitronectin and tube formation in three-dimensional culture. An antiintegrin alphavbeta5 antibody prevented these TAP20 effects. Overexpression of TAP20 also decreased focal adhesion formation in alphavbeta3-deficient cells. The interaction between TAP20 and beta5 integrin cytoplasmic domain was demonstrated by protein coprecipitation and immunoblotting. Thus, the discovery of TAP20, which interacts with integrin beta5 and modulates cell adhesion, migration, and tube formation, further defines a possible pathway to angiogenesis dependent on PKCtheta.

Show MeSH
Related in: MedlinePlus