Flashlight into the Function of Unannotated C11orf52 using Affinity Purification Mass Spectrometry

For an enhanced understanding
of the biological mechanisms of human
disease, it is essential to investigate protein functions. In a previous
study, we developed a prediction method of gene ontology (GO) terms
by the I-TASSER/COFACTOR result, and we applied this to uPE1 in chromosome
11. Here, to validate the bioinformatics prediction of C11orf52, we
utilized affinity purification and mass spectrometry to identify interacting
partners of C11orf52. Using immunoprecipitation methods with three
different peptide tags (Myc, Flag, and 2B8) in HEK 293T cell lines,
we identified 79 candidate proteins that are expected to interact
with C11orf52. The results of a pathway analysis of the GO and STRING
database with candidate proteins showed that C11orf52 could be related
to signaling receptor binding, cell–cell adhesion, and ribosome
biogenesis. Then, we selected three partner candidates of DSG1, JUP,
and PTPN11 for verification of the interaction with C11orf52 and confirmed
them by colocalization at the cell–cell junctions by coimmunofluorescence
experiments. On the basis of this study, we expect that C11orf52 is
related to the Wnt signaling pathway via DSG1 from the protein–protein
interactions, given the results of a comprehensive analysis of the
bioinformatic predictions. The data set is available at the ProteomeXchange
consortium via PRIDE repository (PXD026986).