FHL2 interacts with both ADAM-17 and the cytoskeleton and regulates ADAM-17 localization and activity

J Cell Physiol. 2006 Aug;208(2):363-72. doi: 10.1002/jcp.20671.

Abstract

ADAM-17 is a metalloprotease-disintegrin responsible for the ectodomain shedding of several transmembrane proteins. Using the yeast two-hybrid system, we showed that ADAM-17 interacts with the Four and Half LIM domain 2 protein (FHL2), a LIM domain protein that is involved in multiple protein-protein interaction. We demonstrated that this interaction involved the amino-acid sequence of ADAM-17 from position 721 to739. In the cardiomyoblast cells H9C2, ADAM-17 and FHL2 colocalize with the actin-based cytoskeleton and we showed that FHL2 binds both ADAM-17 and the actin-based cytoskeleton. We found that mainly the mature form of ADAM-17 associates with the cytoskeleton, although the maturation of ADAM-17 by furin is not necessary for its binding to the cytoskeleton. Interestingly, less ADAM-17 was detected at the surface of wild-type mouse macrophages compared to FHL2 deficient macrophages. However, wild-type cells have a higher ability to release ADAM-17 substrates under PMA stimulation. Altogether, these results demonstrate a physical and functional interaction between ADAM-17 and FHL2 that implies that FHL2 has a role in the regulation of ADAM-17.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / chemistry
  • ADAM Proteins / metabolism*
  • ADAM17 Protein
  • Actins / metabolism
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • Cytoskeleton / metabolism*
  • Green Fluorescent Proteins / metabolism
  • Homeodomain Proteins / metabolism*
  • LIM-Homeodomain Proteins
  • Macrophages, Peritoneal / drug effects
  • Macrophages, Peritoneal / metabolism
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Knockout
  • Molecular Sequence Data
  • Muscle Proteins / metabolism*
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / ultrastructure
  • Protein Binding
  • Rats
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transcription Factors / metabolism*

Substances

  • Actins
  • Fhl2 protein, rat
  • Homeodomain Proteins
  • LIM-Homeodomain Proteins
  • Membrane Proteins
  • Muscle Proteins
  • Transcription Factors
  • Green Fluorescent Proteins
  • ADAM Proteins
  • ADAM17 Protein
  • Adam17 protein, mouse
  • Adam17 protein, rat
  • Tetradecanoylphorbol Acetate