Human galectin-16 has a pseudo ligand binding site and plays a role in regulating c-Rel-mediated lymphocyte activity

Yunlong Si, Yuan Yao, Gabriela Jaramillo Ayala, Xumin Li, Qiuyu Han, Wenlu Zhang, Xuejiao Xu, Guihua Tai, Kevin H. Mayo, Yifa Zhou, Jiyong Su

Research output: Contribution to journalArticlepeer-review

Abstract

Background: The structure of human galectin-16 (Gal-16) has yet to be solved, and its function has remained elusive. Methods: X-ray crystallography was used to determine the atomic structures of Gal-16 and two of its mutants. The Gal-16 oligomer state was investigated by gel filtration, its hemagglutination activity was determined along with its ability to bind lactose using ITC. The cellular distribution of EGFP-tagged Gal-16 in various cell lines was also investigated, and the interaction between Gal-16 and c-Rel was assessed by pull-down studies, microscale thermophoresis and immunofluorescence. Results: Unlike other galectins, Gal-16 lacks the ability to bind the β-galactoside lactose. Lactose binding could be regained by replacing an arginine (Arg55) with asparagine, as shown in the crystal structures of two lactose-loaded Gal-16 mutants (R55N and R55N/H57R). Gal-16 was also shown to be monomeric by gel filtration, as well as in crystal structures. Thus, this galectin could not induce erythrocyte agglutination. EGFP-tagged Gal-16 was found to be localized mostly in the nucleus of various cell types, and can interact with c-Rel, a member of NF-κB family. Conclusions: Gal-16 exists as a monomer and its ligand binding is significantly different from that of other prototype galectins, suggesting that it has a novel function(s). The interaction between Gal-16 and c-Rel indicates that Gal-16 may regulate signal transduction pathways via the c-Rel hub in B or T cells at the maternal-fetal interface. General significance: The present study lays the foundation for further studies into the cellular and physiological functions of Gal-16.

Original languageEnglish (US)
Article number129755
JournalBiochimica et Biophysica Acta - General Subjects
Volume1865
Issue number1
DOIs
StatePublished - Jan 2021

Keywords

  • c-Rel
  • Cellular localization
  • Crystal structure
  • Galectin-16
  • Protein-protein interactions

PubMed: MeSH publication types

  • Journal Article

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