1. Academic Validation
  2. Defining the conformational states that enable transglutaminase 2 to promote cancer cell survival versus cell death

Defining the conformational states that enable transglutaminase 2 to promote cancer cell survival versus cell death

  • bioRxiv. 2024 Feb 8:2024.02.04.578794. doi: 10.1101/2024.02.04.578794.
Cody Aplin 1 2 Kara A Zielinski 3 Suzette Pabit 3 Deborah Ogunribido 1 William P Katt 2 Lois Pollack 3 Richard A Cerione 1 2 Shawn K Milano 1 2
Affiliations

Affiliations

  • 1 Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853.
  • 2 Department of Molecular Medicine, Cornell University, Ithaca, NY 14853.
  • 3 School of Applied and Engineering Physics, Cornell University, Ithaca, NY 14853.
Abstract

Transglutaminase 2 (TG2) is a GTP-binding/protein-crosslinking Enzyme that has been investigated as a therapeutic target for Celiac disease, neurological disorders, and aggressive cancers. TG2 has been suggested to adopt two conformational states that regulate its functions: a GTP-bound, closed conformation, and a calcium-bound, crosslinking-active open conformation. TG2 mutants that constitutively adopt an open conformation are cytotoxic to Cancer cells. Thus, small molecules that maintain the open conformation of TG2 could offer a new therapeutic strategy. Here, we investigate TG2, using static and time-resolved small-angle X-ray scattering (SAXS) and single-particle cryoelectron microscopy (cryo-EM), to determine the conformational states responsible for conferring its biological effects. We also describe a newly developed TG2 inhibitor, LM11, that potently kills glioblastoma cells and use SAXS to investigate how LM11 affects the conformational states of TG2. Using SAXS and cryo-EM, we show that guanine nucleotide-bound TG2 adopts a monomeric closed conformation while calcium-bound TG2 assumes an open conformational state that can form higher order oligomers. SAXS analysis also suggests how a TG2 mutant that constitutively adopts the open state binds nucleotides through an alternative mechanism to wildtype TG2. Furthermore, we use time-resolved SAXS to show that LM11 increases the ability of calcium to drive TG2 to an open conformation, which is not reversible by guanine nucleotides and is cytotoxic to Cancer cells. Taken together, our findings demonstrate that the conformational dynamics of TG2 are more complex than previously suggested and highlight how conformational stabilization of TG2 by LM11 maintains TG2 in a cytotoxic conformational state.

Keywords

Biochemistry; Biological Sciences; cancer; cryoelectron microscopy; inhibitors; small angle X-ray scattering; time-resolved; transglutaminase.

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