title: Soluble T cell immunoglobulin and mucin domain (TIM)-1 and -4 generated by A Disintegrin And Metalloprotease (ADAM)-10 and -17 bind to phosphatidylserine.
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.
authors.affiliation: Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Schweigert
authors.affiliation: Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Dewitz
authors.affiliation: Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Möller-Hackbarth
authors.affiliation: Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Trad
authors.affiliation: Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Garbers
authors.affiliation: Institute of Biochemistry, Medical Faculty, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Rose-John
authors.affiliation: Institute of Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University, Düsseldorf, Germany. Electronic address: jscheller@uni-duesseldorf.de.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Scheller
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072197:ADAM10 Protein; D000072198:ADAM17 Protein; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D057809:HEK293 Cells; D000072596:Hepatitis A Virus Cellular Receptor 1; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010718:Phosphatidylserines / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000378:metabolism; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D017384:Sequence Deletion; D012995:Solubility / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000596:Amino Acids; C491143:HAVCR1 protein, human; D000072596:Hepatitis A Virus Cellular Receptor 1; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010718:Phosphatidylserines; D034741:RNA, Small Interfering; D011991:Receptors, Virus; C512411:TIMD4 protein, human; D015759:Ionomycin; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; C491234:Adam10 protein, mouse; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human; C000606463:Adam17 protein, mouse; D013755:Tetradecanoylphorbol Acetate
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