Hanneken A, Maher P A, Baird A
Department of Molecular and Cellular Growth Biology, Whittier Institute for Diabetes and Endocrinology, La Jolla, California 92037.
J Cell Biol. 1995 Mar;128(6):1221-8. doi: 10.1083/jcb.128.6.1221.
We recently characterized three FGF-binding proteins (FGF-BPs) which are soluble forms of the extracellular domains of the high affinity FGF receptors (Hanneken, A. M., W. Ying, N. Ling, and A. Baird. Proc. Natl. Acad. Sci. USA. 1994. 91:9170-9174). These proteins circulate in blood and have been proposed to modulate the biological activity of the FGF family of proteins. Immunohistochemical studies now demonstrate that these soluble, truncated FGF receptors are also present in the basement membranes of retinal vascular endothelial cells. These immunoreactive proteins can be detected with antibodies raised to the extracellular domain of FGFR-1 but not with antibodies raised to either the juxtamembrane domain or the cytoplasmic domain of FGFR-1. Western blotting of human retinal extracts with the antibody raised to the extracellular domain of FGFR-1 detects specific, low molecular mass proteins at 85 kD and 55 kD, corresponding in size to the FGF-BPs, which are not detected with antibodies against the cytoplasmic domain of the receptor. The interaction of this receptor with the extracellular matrix is not dependent on the presence of FGF-2. Immunoreactive receptors are still detected in vascular basement membranes after the removal of FGF-2 with heparitinase. In addition, the recombinant extracellular domain of FGFR-1 continues to bind to corneal endothelial cell matrix after endogenous FGF-2 has been removed with 2 M NaCl. Acid treatment, which has been shown to disrupt protein interactions with the extracellular matrix, leads to a significant reduction in the presence of the matrix form of the FGF receptor. This loss can be restored with exogenous incubations of the recombinant extracellular domain of FGFR-1. This report is the first demonstration that a truncated form of a high affinity growth factor receptor can be localized to the extracellular matrix. These findings add to the list of binding proteins associated with the extracellular matrix (IGFBP-5) and suggest a potentially new regulatory mechanism for controlling the biological availability of FGF, and other peptide growth factors, in the extracellular matrix.
我们最近鉴定了三种FGF结合蛋白(FGF-BPs),它们是高亲和力FGF受体细胞外结构域的可溶性形式(汉内科恩,A.M.,W.英,N.凌,和A.贝尔德。美国国家科学院院刊。1994年。91:9170 - 9174)。这些蛋白在血液中循环,并被认为可调节FGF家族蛋白的生物活性。免疫组织化学研究现在表明,这些可溶性的、截短的FGF受体也存在于视网膜血管内皮细胞的基底膜中。这些免疫反应性蛋白可用针对FGFR-1细胞外结构域产生的抗体检测到,但不能用针对FGFR-1近膜结构域或细胞质结构域产生的抗体检测到。用人视网膜提取物与针对FGFR-1细胞外结构域产生的抗体进行蛋白质印迹分析,可检测到85kD和55kD的特异性低分子量蛋白,其大小与FGF-BPs相对应,而用针对受体细胞质结构域的抗体则检测不到。该受体与细胞外基质的相互作用不依赖于FGF-2的存在。在用肝素酶去除FGF-2后,仍可在血管基底膜中检测到免疫反应性受体。此外,在用2M NaCl去除内源性FGF-2后,FGFR-1的重组细胞外结构域仍继续与角膜内皮细胞基质结合。酸处理已被证明会破坏蛋白质与细胞外基质的相互作用,导致FGF受体基质形式的存在显著减少。这种损失可通过FGFR-1重组细胞外结构域的外源孵育来恢复。本报告首次证明高亲和力生长因子受体的截短形式可定位于细胞外基质。这些发现增加了与细胞外基质相关的结合蛋白(IGFBP-5)的种类,并提示了一种潜在的新调控机制,用于控制细胞外基质中FGF和其他肽生长因子的生物可利用性。