Impaired activity and membrane association of most calpain-5 mutants causal for neovascular inflammatory vitreoretinopathy

James W. Geddes, Vimala Bondada, Dorothy E. Croall, David W. Rodgers, Jozsef Gal

Research output: Contribution to journalArticlepeer-review


Neovascular inflammatory vitreoretinopathy (NIV) is a rare eye disease that ultimately leads to complete blindness and is caused by mutations in the gene encoding calpain-5 (CAPN5), with six pathogenic mutations identified. In transfected SH-SY5Y cells, five of the mutations resulted in decreased membrane association, diminished S-acylation, and reduced calcium-induced autoproteolysis of CAPN5. CAPN5 proteolysis of the autoimmune regulator AIRE was impacted by several NIV mutations. R243, L244, K250 and the adjacent V249 are on β-strands in the protease core 2 domain. Conformational changes induced by Ca2+binding result in these β-strands forming a β-sheet and a hydrophobic pocket which docks W286 side chain away from the catalytic cleft, enabling calpain activation based on comparison with the Ca2+-bound CAPN1 protease core. The pathologic variants R243L, L244P, K250N, and R289W are predicted to disrupt the β-strands, β-sheet, and hydrophobic pocket, impairing calpain activation. The mechanism by which these variants impair membrane association is unclear. G376S impacts a conserved residue in the CBSW domain and is predicted to disrupt a loop containing acidic residues which may contribute to membrane binding. G267S did not impair membrane association and resulted in a slight but significant increase in autoproteolytic and proteolytic activity. However, G267S is also identified in individuals without NIV. Combined with the autosomal dominant pattern of NIV inheritance and evidence that CAPN5 may dimerize, the results are consistent with a dominant negative mechanism for the five pathogenic variants which resulted in impaired CAPN5 activity and membrane association and a gain-of-function for the G267S variant.

Original languageEnglish
Article number166747
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Issue number6
StatePublished - Aug 2023

Bibliographical note

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© 2023 The Authors


  • Autoproteolysis
  • CAPN5
  • Calpain-5
  • Membrane
  • NIV
  • Palmitoylation
  • S-acylation

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology


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