A transmembrane tight junction protein selectively expressed on endothelial cells and platelets

被引:162
作者
Nasdala, I
Wolburg-Buchholz, K
Wolburg, H
Kuhn, A
Ebnet, K
Brachtendorf, G
Samulowitz, U
Kuster, B
Engelhardt, B
Vestweber, D
Butz, S
机构
[1] Univ Munster, ZMBE, Inst Cell Biol, D-48149 Munster, Germany
[2] Max Planck Inst, D-69117 Heidelberg, Germany
[3] Cellzome AG, D-72076 Tubingen, Germany
[4] Univ Tubingen, Inst Pathol, D-7400 Tubingen, Germany
关键词
D O I
10.1074/jbc.M111999200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Searching for cell surface proteins expressed at interendothelial cell contacts, we have raised monoclonal antibodies against intact mouse endothelial cells. We obtained two monoclonal antibodies, 1G8 and 4C10, that stain endothelial cell contacts and recognize a protein of 55 kDa. Purification and identification by mass spectrometry of this protein revealed that it contains two extracellular Ig domains, reminiscent of the JAM family, but a much longer 120-amino acid cytoplasmic domain. The antigen is exclusively expressed on endothelial cells of various organs as was analyzed by immunohistochemistry. Immunogold labeling of ultrathin sections of brain as well as skeletal muscle revealed that the antigen strictly colocalizes in capillaries with the tight junction markers occludin, claudin-5, and ZO-1. Upon transfection into MDCK cells, the antigen was restricted to the most apical tip of the lateral cell surface, where it colocalized with ZO-1 but not with beta-catenin. In contrast to JAM-1, however, the 1G8 antigen does not associate with the PDZ domain proteins ZO-1, AF-6, or ASIP/PAR-3, despite the presence of a PDZ-binding motif. The 1G8 antigen was not detected on peripheral blood mouse leukocytes, whereas similar to JAM-1 it was strongly expressed on platelets and megakaryocytes. The 1G8 antigen supports homophilic interactions on transfected Chinese hamster ovary cells. Based on the similarity to the JAM molecules, it is plausible that the 1G8 antigen might be involved in interendothelial cell adhesion.
引用
收藏
页码:16294 / 16303
页数:10
相关论文
共 46 条
  • [1] ANDERSON JM, 1995, AM J PHYSIOL, V269, P467
  • [2] Cloning of human junctional adhesion molecule 3 (JAM3) and its identification as the JAM2 counter-receptor
    Arrate, MP
    Rodriguez, JM
    Tran, TM
    Brock, TA
    Cunningham, SA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (49) : 45826 - 45832
  • [3] JAM-2, a novel immunoglobulin superfamily molecule, expressed by endothelial and lymphatic cells
    Aurrand-Lions, M
    Duncan, L
    Ballestrem, C
    Imhof, BA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (04) : 2733 - 2741
  • [4] Aurrand-Lions MA, 2000, CURR TOP MICROBIOL, V251, P91
  • [5] Chrétien I, 1998, EUR J IMMUNOL, V28, P4094
  • [6] Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo
    Corada, M
    Mariotti, M
    Thurston, G
    Smith, K
    Kunkel, R
    Brockhaus, M
    Lampugnani, MG
    Martin-Padura, I
    Stoppacciaro, A
    Ruco, L
    McDonald, DM
    Ward, PA
    Dejana, E
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (17) : 9815 - 9820
  • [7] A novel protein with homology to the junctional adhesion molecule - Characterization of leukocyte interactions
    Cunningham, SA
    Arrate, MP
    Rodriguez, JM
    Bjercke, RJ
    Vanderslice, P
    Morris, AP
    Brock, TA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (44) : 34750 - 34756
  • [8] Dejana E, 2000, INT J DEV BIOL, V44, P743
  • [9] Leukocyte recruitment in the cerebrospinal fluid of mice with experimental meningitis is inhibited by an antibody to junctional adhesion molecule (JAM)
    Del Maschio, A
    De Luigi, A
    Martin-Padura, I
    Brockhaus, M
    Bartfai, T
    Fruscella, P
    Adorini, L
    Martino, GV
    Furlan, R
    De Simoni, MG
    Dejana, E
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1999, 190 (09) : 1351 - 1356
  • [10] Ebnet K, 2000, J BIOL CHEM, V275, P27979