Galectin-3 bound cell membrane proteins identified by LC-MS/MS

The HMEC-1 membrane proteins were divided into two fractions: one for His-Gal-3 with a His-tag antibody, and the other for the His-tag antibody alone. Subsequently, SDS-PAGE and silver staining were performed. Mass spectrometry was conducted to analyze potential Gal-3 binding candidates, resulting in the identification of a total of 64 distinct proteins. Given that Gal-3 is believed to bind to cell surface glycoproteins, thereby mediating various functions, the selected proteins were those with the cell membrane. Among these, 15 proteins were primarily localized in the cell membrane, each with unique peptides exceeding two. In our study, we found that the top five proteins bound with Gal-3 identified in HMEC-1 were CD166, CD146, ECE1, integrin β1, and catenin α-1. Taken together, this study identifies the candidate proteins bound to Gal-3 and suggest that the proteins binding to Gal-3 vary on the cell types from different tissues.