Ostman A, Yang Q, Tonks N K
Cold Spring Harbor Laboratory, NY 11724-2208.
Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9680-4. doi: 10.1073/pnas.91.21.9680.
cDNA encoding a receptor-like protein-tyrosine-phosphatase (PTP) termed DEP-1 was isolated from a HeLa cell library. The cDNA predicts an enzyme consisting of an extracellular segment containing eight fibronectin type III repeats, a single transmembrane segment, and a single intracellular PTP domain. Following expression of DEP-1 cDNA in COS cells a glycoprotein of 180 kDa was detected and PTP activity was demonstrated in immunocomplexes with a C-terminal peptide antiserum. Endogenous DEP-1 was detected in WI-38 human embryonic lung fibroblasts by immunoblotting and immunocomplex PTP assays. Immunoblot analysis of DEP-1 expression in WI-38 cells revealed dramatically increased levels and activity of the PTP in dense cultures relative to sparse cultures. Also, DEP-1 activity, detected in PTP assays of immunocomplexes, was increased in dense cell cultures. In contrast, the expression levels of PTP-1B did not change with cell density. This enhancement of DEP-1 expression with increasing cell density was also observed in another fibroblast cell line, AG1518. The increase in DEP-1 occurs gradually with increasing cell contact and is initiated before saturation cell density is reached. These observations suggest that DEP-1 may contribute to the mechanism of contact inhibition of cell growth.
从HeLa细胞文库中分离出编码一种名为DEP-1的受体样蛋白酪氨酸磷酸酶(PTP)的cDNA。该cDNA预测的一种酶由包含八个III型纤连蛋白重复序列的细胞外区段、单个跨膜区段和单个细胞内PTP结构域组成。在COS细胞中表达DEP-1 cDNA后,检测到一种180 kDa的糖蛋白,并在与C末端肽抗血清形成的免疫复合物中证实了PTP活性。通过免疫印迹和免疫复合物PTP分析在WI-38人胚肺成纤维细胞中检测到内源性DEP-1。对WI-38细胞中DEP-1表达的免疫印迹分析显示,相对于稀疏培养,在致密培养中PTP的水平和活性显著增加。此外,在免疫复合物的PTP分析中检测到的DEP-1活性在致密细胞培养中增加。相比之下,PTP-1B的表达水平不随细胞密度而变化。在另一种成纤维细胞系AG1518中也观察到随着细胞密度增加DEP-1表达增强。DEP-1的增加随着细胞接触增加而逐渐发生,并在达到饱和细胞密度之前就开始了。这些观察结果表明DEP-1可能参与细胞生长接触抑制的机制。