The Biophysical Interaction of the Danger-Associated Molecular Pattern (DAMP) Calreticulin with the Pattern-Associated Molecular Pattern (PAMP) Lipopolysaccharide

Int J Mol Sci. 2019 Jan 18;20(2):408. doi: 10.3390/ijms20020408.

Abstract

The endoplasmic reticulum (ER) chaperone protein, calreticulin (CRT), is essential for proper glycoprotein folding and maintaining cellular calcium homeostasis. During ER stress, CRT is overexpressed as part of the unfolded protein response (UPR). In addition, CRT can be released as a damage-associated molecular pattern (DAMP) molecule that may interact with pathogen-associated molecular patterns (PAMPs) during the innate immune response. One such PAMP is lipopolysaccharide (LPS), a component of the gram-negative bacterial cell wall. In this report, we show that recombinant and native human placental CRT strongly interacts with LPS in solution, solid phase, and the surface of gram-negative and gram-positive bacteria. Furthermore, LPS induces oilgomerization of CRT with a disappearance of the monomeric form. The application of recombinant CRT (rCRT) to size exclusion and anion exchange chromatography shows an atypical heterogeneous elution profile, indicating that LPS affects the conformation and ionic charge of CRT. Interestingly, LPS bound to CRT is detected in sera of bronchiectasis patients with chronic bacterial infections. By ELISA, rCRT dose-dependently bound to solid phase LPS via the N- and C-domain globular head region of CRT and the C-domain alone. The specific interaction of CRT with LPS may be important in PAMP innate immunity.

Keywords: Danger Associated Molecular Patterns (DAMPs); Pathogen Associated Molecular Patterns (PAMPs); bacterial opsonin; calreticulin; endoplasmic reticulum chaperone; innate immunity; lipopolysaccharide.

MeSH terms

  • Alarmins / chemistry
  • Alarmins / metabolism*
  • Animals
  • Calreticulin / chemistry
  • Calreticulin / metabolism*
  • Chromatography, Gel
  • Endotoxins / metabolism
  • Humans
  • Lipopolysaccharides / metabolism*
  • Polysaccharides, Bacterial / chemistry
  • Polysaccharides, Bacterial / metabolism
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Multimerization
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Alarmins
  • Calreticulin
  • Endotoxins
  • Lipopolysaccharides
  • Polysaccharides, Bacterial
  • Recombinant Proteins