Substrate-selective protein ectodomain shedding by ADAM17 and iRhom2 depends on their juxtamembrane and transmembrane domains

FASEB J. 2020 Apr;34(4):4956-4969. doi: 10.1096/fj.201902649R. Epub 2020 Feb 26.

Abstract

The metalloprotease ADAM17 (a disintegrin and metalloprotease 17) regulates EGF-receptor and TNFα signaling, thereby not only protecting the skin and intestinal barrier, but also contributing to autoimmunity. ADAM17 can be rapidly activated by many stimuli through its transmembrane domain (TMD), with the seven membrane-spanning inactive Rhomboids (iRhom) 1 and 2 implicated as candidate regulatory partners. However, several alternative models of ADAM17 regulation exist that do not involve the iRhoms, such as regulation through disulfide bond exchange or through interaction with charged phospholipids. Here, we report that a non-activatable mutant of ADAM17 with the TMD of betacellulin (BTC) can be rescued by restoring residues from the ADAM17 TMD, but only in Adam17-/- cells, which contain iRhoms, not in iRhom1/2-/- cells. We also provide the first evidence that the extracellular juxtamembrane domains (JMDs) of ADAM17 and iRhom2 regulate the stimulation and substrate selectivity of ADAM17. Interestingly, a point mutation in the ADAM17 JMD identified in a patient with Tetralogy of Fallot, a serious heart valve defect, affects the substrate selectivity of ADAM17 toward Heparin-binding epidermal growth factor like growth factor (HB-EGF), a crucial regulator of heart valve development in mice. These findings provide new insights into the regulation of ADAM17 through an essential interaction with the TMD1 and JMD1 of iRhom2.

Keywords: ADAM17 (a disintegrin and metalloprotease 17); EGFR (epidermal growth factor receptor); Rhbdf1/2 (rhomboid family member 1/2); TGFα (transforming growth factor α); TNFα (tumor necrosis factor α); iRhom1/2 (inactive Rhomboid like protein 1/2).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • ADAM17 Protein / chemistry
  • ADAM17 Protein / genetics
  • ADAM17 Protein / metabolism*
  • Animals
  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cells, Cultured
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mutation*
  • Protein Domains
  • Substrate Specificity
  • Tetralogy of Fallot / genetics

Substances

  • Carrier Proteins
  • iRhom2 protein, mouse
  • ADAM17 Protein
  • Adam17 protein, mouse