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Molecular and Cellular Biology, October 2004, p. 9198-9206, Vol. 24, No. 20
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.20.9198-9206.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Endothelial Cell-Specific Antibody PAL-E Identifies a Secreted Form of Vimentin in the Blood Vasculature

Bin Xu,1 Robert M. deWaal,2 Nirit Mor-Vaknin,3 Chris Hibbard,1 David M. Markovitz,3 and Mark L. Kahn1*

Division of Cardiology, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania,1 Department of Pathology, University Medical Center Nijmegen, Nijmegen, The Netherlands,2 Department of Internal Medicine, Division of Infectious Diseases, University of Michigan, Ann Arbor, Michigan3

Received 28 January 2004/ Returned for modification 23 February 2004/ Accepted 21 July 2004

During mammalian vascular development, endothelial cells form a complex array of vessels that differ markedly in structure and function, but the molecular basis for this vascular complexity is poorly understood. Recent insights into endothelial diversity have come from the identification of molecular markers expressed on distinct endothelial cell populations. One such marker, the PAL-E antibody, has been used for almost 20 years to distinguish blood and lymphatic vessels, but the identity of the protein recognized by PAL-E has been unknown. In the present study we have used protein purification and tandem mass spectrometry analysis of tryptic peptides to identify the PAL-E antigen as a secreted form of vimentin. Vimentin has been well characterized as an intracellular intermediate filament protein expressed broadly in mesenchymal cells. In contrast, PAL-E-reactive vimentin is secreted extracellularly, its synthesis is restricted to a distinct population of blood endothelial cells and activated macrophages, and PAL-E-reactive vimentin is found in circulating human blood. PAL-E-reactive vimentin does not arise from an endothelial cell-specific mRNA transcript but is the product of cell-specific posttranslational modification. The PAL-E antibody therefore defines secretion of vimentin as a molecular distinction among endothelial cells and exposes a novel, extracellular role for vimentin in the blood vasculature.

* Corresponding author. Mailing address: University of Pennsylvania, 421 Curie Blvd., BRB II/III Room 952, Philadelphia, PA 19104-6100. Phone: (215) 898-9007. Fax: (215) 573-2094. E-mail: markkahn{at}mail.med.upenn.edu.

Molecular and Cellular Biology, October 2004, p. 9198-9206, Vol. 24, No. 20
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.20.9198-9206.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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