Expression Cloning of Putative Mouse EF-L1 Receptor
Date Issued
2010
Date
2010
Author(s)
An, Kuan-Yu
Abstract
EF-L1 (epithelial cell produced factor, clone L1; also named proline rich acidic protein 1, PRAP1) is a secreted protein expressed in mouse intestinal epithelial cells (IECs). Our earlier study demonstrated that deficiency of EF-L1 resulted in precocious apoptosis of the differentiated epithelial cells in the villus. Activation of PI3K-Akt pathway was impaired in EF-L1 deficient IECs; oral feeding of recombinant EF-L1 could rescue the phenotype of EF-L1 deficient IECs, suggesting that EF-L1 might function via an autocrine mechanism to affect IECs survival. Moreover, EF-L1 bound to isolated IECs in a specific manner, suggesting that there could be a specific interacting protein or a putative EF-L1 receptor residing on the surface of IECs. We thus constructed a mouse IEC cDNA library and screened for the putative EF-L1 receptor by an expression cloning approach. After quaternary screening, we narrowed down the target pools from ~25000 clones to ~80 clones per pool. Two membrane proteins have been identified in this candidate pool, i.e., transmembrane protein 160 (Tmem160) and the ABC transporter, Abcb1a (ATP-binding cassette, sub-family B, member 1A). Further experiments are needed to confirm the interaction between EF-L1 and the above candidates.
Subjects
expression cloning
intestinal epithilial cell
Type
thesis
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