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© The Rockefeller University Press, 0021-9525/1998//791 $5.00
The Journal of Cell Biology, Volume 141, Number 3, , 1998 791-804

Regulation of Endothelial Cell Motility by Complexes of Tetraspan Molecules CD81/TAPA-1 and CD151/PETA-3 with 3β1 Integrin Localized at Endothelial Lateral Junctions

María Yáñez-Mó^*, Arántzazu Alfranca^*, Carlos Cabañas, Mónica Marazuela, Reyes Tejedor^*, M. Angeles Ursa^*, Leonie K. Ashman^||, Manuel O. de Landázuri^*, and Francisco Sánchez-Madrid^*

^* Servicio de Inmunología and Servicio de Endocrinología, Hospital de la Princesa, Universidad Autónoma de Madrid; Departamento de Bioquímica, Facultad de Medicina, Universidad Complutense, Madrid, Spain; and ^|| Division of Haematology, Institute of Medical and Veterinary Science, University of Adelaide, SA 5000, Australia

Cell-to-cell junction structures play a key role in cell growth rate control and cell polarization. In endothelial cells (EC), these structures are also involved in regulation of vascular permeability and leukocyte extravasation. To identify novel components in EC intercellular junctions, mAbs against these cells were produced and selected using a morphological screening by immunofluorescence microscopy. Two novel mAbs, LIA1/1 and VJ1/16, specifically recognized a 25-kD protein that was selectively localized at cell–cell junctions of EC, both in the primary formation of cell monolayers and when EC reorganized in the process of wound healing. This antigen corresponded to the recently cloned platelet-endothelial tetraspan antigen CD151/PETA-3 (platelet-endothelial tetraspan antigen-3), and was consistently detected at EC cell–cell contact sites. In addition to CD151/PETA-3, two other members of the tetraspan superfamily, CD9 and CD81/ TAPA-1 (target of antiproliferative antibody-1), localized at endothelial cell-to-cell junctions. Biochemical analysis demonstrated molecular associations among tetraspan molecules themselves and those of CD151/ PETA-3 and CD9 with 3β1 integrin. Interestingly, mAbs directed to both CD151/PETA-3 and CD81/ TAPA-1 as well as mAb specific for 3 integrin, were able to inhibit the migration of ECs in the process of wound healing. The engagement of CD151/PETA-3 and CD81/TAPA-1 inhibited the movement of individual ECs, as determined by quantitative time-lapse video microscopy studies. Furthermore, mAbs against the CD151/PETA-3 molecule diminished the rate of EC invasion into collagen gels. In addition, these mAbs were able to increase the adhesion of EC to extracellular matrix proteins. Together these results indicate that CD81/TAPA-1 and CD151/PETA-3 tetraspan molecules are components of the endothelial lateral junctions implicated in the regulation of cell motility, either directly or by modulation of the function of the associated integrin heterodimers.

Abbreviations used in this paper: EC, endothelial cells; ECM, extracellular matrix; HMEC-1, human microvascular endothelial cell line-1; HUVEC, human EC from umbilical vein; PECAM, platelet-endothelial cell adhesion molecule; TM4, transmembrane 4/tetraspan molecule; TNF-, tumor necrosis factor-; PETA-3, platelet-endothelial tetraspan antigen-3; TAPA-1, target of antiproliferative antibody-1;VE, vascular endothelial.

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