Physical tuning of galectin-3 signaling

Proc Natl Acad Sci U S A. 2021 May 11;118(19):e2024117118. doi: 10.1073/pnas.2024117118.

Abstract

Galectin-3 (Gal3) exhibits dynamic oligomerization and promiscuous binding, which can lead to concomitant activation of synergistic, antagonistic, or noncooperative signaling pathways that alter cell behavior. Conferring signaling pathway selectivity through mutations in the Gal3-glycan binding interface is challenged by the abundance of common carbohydrate types found on many membrane glycoproteins. Here, employing alpha-helical coiled-coils as scaffolds to create synthetic Gal3 constructs with defined valency, we demonstrate that oligomerization can physically regulate extracellular signaling activity of Gal3. Constructs with 2 to 6 Gal3 subunits ("Dimer," "Trimer," "Tetramer," "Pentamer," "Hexamer") demonstrated glycan-binding properties and cell death-inducing potency that scaled with valency. Dimer was the minimum functional valency. Unlike wild-type Gal3, which signals apoptosis and mediates agglutination, synthetic Gal3 constructs induced cell death without agglutination. In the presence of CD45, Hexamer was distributed on the cell membrane, whereas it clustered in absence of CD45 via membrane glycans other than those found on CD7. Wild-type Gal3, Pentamer, and Hexamer required CD45 and CD7 to signal apoptosis, and the involvement of caspases in apoptogenic signaling was increased in absence of CD45. However, wild-type Gal3 depended on caspases to signal apoptosis to a greater extent than Hexamer, which had greater caspase dependence than Pentamer. Diminished caspase activation downstream of Hexamer signaling led to decreased pannexin-1 hemichannel opening and interleukin-2 secretion, events facilitated by the increased caspase activation downstream of wild-type Gal3 signaling. Thus, synthetic fixation of Gal3 multivalency can impart physical control of its outside-in signaling activity by governing membrane glycoprotein engagement and, in turn, intracellular pathway activation.

Keywords: coiled-coil; emergent function; galectin-3; multivalency; signal transduction.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics*
  • Blood Proteins / chemistry
  • Blood Proteins / genetics*
  • Blood Proteins / metabolism
  • Cell Death / genetics
  • Cell Line, Tumor
  • Galectins / chemistry
  • Galectins / genetics*
  • Galectins / metabolism
  • Humans
  • Jurkat Cells
  • Lactose / metabolism
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / metabolism
  • Microscopy, Confocal
  • Polysaccharides / metabolism
  • Protein Binding
  • Protein Multimerization
  • Signal Transduction / genetics*
  • T-Lymphocytes / metabolism*

Substances

  • Blood Proteins
  • Galectins
  • LGALS3 protein, human
  • Polysaccharides
  • Leukocyte Common Antigens
  • Lactose