Screening For Candidate Molecule Interacts With LINGO-1 In Yeast Two Hybrid System

Intend to find out what serves as the signaling conductor downstream of LINGO-1, we adopted a GAL-4 Based Two Hybrid Screening system, with the introplasma domain of LINGO-1 as bait, and screened in a human fetal brain cDNA library. After discriminating the most false-positive clones by positive and negative transcriptional activity detection, we finally pick out WNK1 as a candidate. And we considered WNK3 as another candidate in the further study. Then we found that full length LINGO-1 and WNK1(123-510aa) can interact with each other in living COS-7 by FRET detection .By using immunohistochemistry methods, we demonstrated that LINGO-1, WNKland WNK3 are all expressed in β III Tubulin positive neuron and 04 positive oligodendrocyte precursors, thus they maybe co-localized in neurons and oligodendrocyte. Coimmunoprecipitation of endogenous proteins also demonstrate LINGO-1 and WNK1 or WNK3 are combined in neuronal system. Based on these discoveries, we propose that LINGO-1 can interact with WNK1 or WNK3, and it brings a new insight into the biological function of LINGO-1.