ABSTRACT
Ephrins are cell membrane-associated signaling proteins bound by transmembrane Eph receptors on juxtaposed cells. Eph-ephrin interactions result in bidirectional signaling within both receptor- and ligand-bearing cells, with diverse consequences for cell morphology and behavior. Such interactions are especially important during early vertebrate development, and growing evidence has revealed equally important roles in adult-tissue homeostasis. As for the Eph receptors, abnormal expression of ephrins is associated with disease, especially cancer. The ephrins have received less attention than the Ephs in the literature, owing, in part, to their later discovery and that they are fewer in number. Here, we attempt to redress this imbalance and provide an 'ephrin-centric' discussion of the expression and function of ephrins in cancer.
Subject(s)
Ephrins/metabolism , Neoplasms/metabolism , Receptors, Eph Family/metabolism , Signal Transduction/physiology , Animals , HumansABSTRACT
Eph receptor tyrosine kinases (RTKs) are a highly conserved family of signaling proteins with functions in cellular migration, adhesion, apoptosis, and proliferation during both adult and embryonic life. Here, we describe a knock-in mouse in which EphA1 expression is disrupted via the insertion of an internal ribosome entry site (IRES)-human placental alkaline phosphatase (ALPP) reporter cassette into exon II of the EphA1 gene. This was shown to successfully knockout expression of endogenous EphA1 and enforce expression of the ALPP reporter by the EphA1 promoter. Staining for the ALPP reporter protein demonstrated an epithelially restricted expression pattern in mouse tissues. In EphA1 null mice, two separate phenotypes were identified: abnormal tail development manifesting as a kinky tail was found in approximately 80% of homozygous adults. A second, distinct abnormality present in approximately 18% of females was characterized by imperforate uterovaginal development with hydrometrocolpos and caused by a resistance of cells to apoptosis during reproductive tract canalization. These results indicate a possible role for EphA1 in tissue patterning and hormone-induced apoptotic processes.
Subject(s)
Genes, Reporter , Receptor, EphA1/genetics , Alkaline Phosphatase , Animals , Apoptosis/genetics , Body Patterning/genetics , Ephrin-A1/metabolism , Female , GPI-Linked Proteins , Gene Knock-In Techniques , Humans , Isoenzymes/genetics , Male , Mice , Mice, Knockout , Receptor, EphA1/physiology , Tail/abnormalities , Tail/cytology , Tail/enzymology , Uterus/abnormalities , Uterus/cytology , Uterus/enzymology , Vagina/abnormalities , Vagina/cytology , Vagina/enzymologyABSTRACT
There is an urgent need for high purity, single chain, fully functional Eph/ephrin membrane proteins. This report outlines the pTIg-BOS-Fc vector and purification approach resulting in rapid increased production of fully functional single chain extracellular proteins that were isolated with high purity and used in structure-function analysis and pre-clinical studies.
