Soluble T cell immunoglobulin and mucin domain (TIM)-1 and -4 generated by A Disintegrin And Metalloprotease (ADAM)-10 and -17 bind to phosphatidylserine

Olga Schweigert; Christin Dewitz; Katja Möller-Hackbarth; Ahmad Trad; Christoph Garbers; Stefan Rose-John; Jürgen Scheller

doi:10.1016/j.bbamcr.2013.11.014

Soluble T cell immunoglobulin and mucin domain (TIM)-1 and -4 generated by A Disintegrin And Metalloprotease (ADAM)-10 and -17 bind to phosphatidylserine

Biochim Biophys Acta. 2014 Feb;1843(2):275-87. doi: 10.1016/j.bbamcr.2013.11.014. Epub 2013 Nov 25.

Authors

Olga Schweigert¹, Christin Dewitz¹, Katja Möller-Hackbarth¹, Ahmad Trad¹, Christoph Garbers², Stefan Rose-John¹, Jürgen Scheller³

Affiliations

¹ Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
² Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, Germany.
³ Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, Germany. Electronic address: jscheller@uni-duesseldorf.de.

PMID: 24286866
DOI: 10.1016/j.bbamcr.2013.11.014

Abstract

T cell immunoglobulin and mucin domain 1 and 4 (TIM-1 and -4) proteins serve as phosphatidylserine receptors to engulf apoptotic cells. Here we show that human TIM-1 and TIM-4 proteins are targets of A Disintegrin And Metalloprotease (ADAM)-mediated ectodomain shedding resulting in soluble forms of TIM-1 and TIM-4. We identified ADAM10 and ADAM17 as major sheddases of TIM-1 and TIM-4 as shown by protease-specific inhibitors, the ADAM10 prodomain, siRNA and ADAM10/ADAM17 deficient murine embryonic fibroblasts (MEFs). TIM-1 and TIM-4 lacking the intracellular domain were efficiently cleaved after ionomycin- and PMA-treatment, indicating that the intracellular domain was not necessary for ectodomain shedding. Soluble TIM-1 and -4 were able to bind to phosphatidylserine, suggesting that soluble TIM-1 and -4 might act as negative regulators of cellular TIM-1 and -4. In summary, we describe TIM-1 and TIM-4 as novel targets for ADAM10- and ADAM17-mediated ectodomain shedding.

Keywords: ADAM; Cell surface receptor; DMEM; Dulbecco's modified Eagle's medium; FBS; Shedding; T cell immunoglobulin and mucin domain protein; fetal bovine serum; mAb; monoclonal antibody.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADAM Proteins / metabolism*
ADAM10 Protein
ADAM17 Protein
Amino Acid Sequence
Amino Acids / metabolism
Amyloid Precursor Protein Secretases / metabolism*
Animals
Cell Differentiation / drug effects
Cell Membrane / drug effects
Cell Membrane / metabolism
HEK293 Cells
Hepatitis A Virus Cellular Receptor 1
Humans
Ionomycin / pharmacology
Membrane Glycoproteins / chemistry
Membrane Glycoproteins / metabolism*
Membrane Proteins / chemistry
Membrane Proteins / metabolism*
Mice
Molecular Sequence Data
Phosphatidylserines / metabolism*
Protein Binding / drug effects
Protein Structure, Tertiary
RNA, Small Interfering / metabolism
Receptors, Virus / chemistry
Receptors, Virus / metabolism*
Sequence Deletion
Solubility / drug effects
Tetradecanoylphorbol Acetate / pharmacology

Substances

Amino Acids
HAVCR1 protein, human
Hepatitis A Virus Cellular Receptor 1
Membrane Glycoproteins
Membrane Proteins
Phosphatidylserines
RNA, Small Interfering
Receptors, Virus
TIMD4 protein, human
Ionomycin
Amyloid Precursor Protein Secretases
ADAM Proteins
ADAM10 Protein
ADAM10 protein, human
Adam10 protein, mouse
ADAM17 Protein
ADAM17 protein, human
Adam17 protein, mouse
Tetradecanoylphorbol Acetate