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1
* Zentrum Anatomie, Abteilung Histologie, Universität, Göttingen, D-37075 Göttingen, Germany; and
Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany
1Correspondence: Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, D-82152 Martinsried, Germany. E-mail: TIMPL{at}biochem.mpg.de
Theendothelial cell inhibitor endostatin (22 kDa) is part of the carboxyl-terminal globular domain of collagen XVIII and shows a widespread tissue distribution. Immunohistology of adult mouse tissues demonstrated a preferred localization in many vessel walls and some other basement membrane zones. A strong immunogold staining was observed across elastic fibers in the multiple elastic membranes of aorta and other large arteries. Staining was less strong along sparse elastic fibers of veins and almost none was observed in the walls of arterioles and capillaries. Strong evidence was also obtained for some intracellular and basement membrane associations. Immunogold double staining of elastic fibers showed a close colocalization of endostatin with fibulin-2, fibulin-1, and nidogen-2, but not with perlecan. Reasonable amounts of endostatin could be extracted from aorta and skin by EDTA, followed by detergents, with aorta being the richest source of the inhibitor identified so far. Solubilizations with collagenase and elastase were ~fivefold less efficient. Immunoblots of aortic extracts detected major endostatin components of 22–25 kDa whereas skin extracts also contained some larger components. Solid-phase assays demonstrated distinct binding of recombinant mouse endostatin to the fibulins and nidogen-2, consistent with their tissue colocalization. Together, the data indicate several different ways for endostatin to be associated with the extracellular matrix, and its release may determine biological activation. This also defines a novel function for some elastic tissues.—Miosge, N., Sasaki, T., Timpl, R. Angiogenesis inhibitor endostatin is a distinct component of elastic fibers in vessel walls.
Key Words: binding assays • extracellular matrix • immunogold staining
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