Cell-surface metalloprotease ADAM12 is internalized by a clathrin- and Grb2-dependent mechanism
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Cell-surface metalloprotease ADAM12 is internalized by a clathrin- and Grb2-dependent mechanism. / Hansen, Dorte Stautz; Leyme, Anthony; Grandal, Michael Vibo; Albrechtsen, Reidar; van Deurs, Bo; Wewer, Ulla; Kveiborg, Marie.
In: Traffic - International Journal of Intracellular Transport, Vol. 13, No. 11, 2012, p. 1532-46.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Cell-surface metalloprotease ADAM12 is internalized by a clathrin- and Grb2-dependent mechanism
AU - Hansen, Dorte Stautz
AU - Leyme, Anthony
AU - Grandal, Michael Vibo
AU - Albrechtsen, Reidar
AU - van Deurs, Bo
AU - Wewer, Ulla
AU - Kveiborg, Marie
N1 - © 2012 John Wiley & Sons A/S.
PY - 2012
Y1 - 2012
N2 - ADAM12 (A Disintegrin And Metalloprotease 12), a member of the ADAMs family of transmembrane proteins, is involved in ectodomain shedding, cell-adhesion and signaling, with important implications in cancer. Therefore, mechanisms that regulate the levels and activity of ADAM12 at the cell-surface are possibly crucial in these contexts. We here investigated internalization and subsequent recycling or degradation of ADAM12 as a potentially important regulatory mechanism. Our results show that ADAM12 is constitutively internalized primarily via the clathrin-dependent pathway and is subsequently detected in both early and recycling endosomes. The protease activity of ADAM12 does not influence this internalization mechanism. Analysis of essential elements for internalization established that proline-rich regions in the cytoplasmic domain of ADAM12, previously shown to interact with Src-homology 3 domains, were necessary for proper internalization. These sites in the ADAM12 cytoplasmic domain interacted with the adaptor protein growth factor receptor-bound protein 2 (Grb2) and knockdown of Grb2 markedly reduced ADAM12 internalization. These studies establish that internalization is indeed a mechanism that regulates ADAM cell surface levels and show that ADAM12 internalization involves the clathrin-dependent pathway and Grb2.
AB - ADAM12 (A Disintegrin And Metalloprotease 12), a member of the ADAMs family of transmembrane proteins, is involved in ectodomain shedding, cell-adhesion and signaling, with important implications in cancer. Therefore, mechanisms that regulate the levels and activity of ADAM12 at the cell-surface are possibly crucial in these contexts. We here investigated internalization and subsequent recycling or degradation of ADAM12 as a potentially important regulatory mechanism. Our results show that ADAM12 is constitutively internalized primarily via the clathrin-dependent pathway and is subsequently detected in both early and recycling endosomes. The protease activity of ADAM12 does not influence this internalization mechanism. Analysis of essential elements for internalization established that proline-rich regions in the cytoplasmic domain of ADAM12, previously shown to interact with Src-homology 3 domains, were necessary for proper internalization. These sites in the ADAM12 cytoplasmic domain interacted with the adaptor protein growth factor receptor-bound protein 2 (Grb2) and knockdown of Grb2 markedly reduced ADAM12 internalization. These studies establish that internalization is indeed a mechanism that regulates ADAM cell surface levels and show that ADAM12 internalization involves the clathrin-dependent pathway and Grb2.
KW - ADAM Proteins
KW - Breast Neoplasms
KW - Carcinoma
KW - Clathrin
KW - Endocytosis
KW - Endosomes
KW - Female
KW - GRB2 Adaptor Protein
KW - HEK293 Cells
KW - Humans
KW - Membrane Proteins
KW - Proline-Rich Protein Domains
KW - Protein Interaction Domains and Motifs
U2 - 10.1111/j.1600-0854.2012.01405.x
DO - 10.1111/j.1600-0854.2012.01405.x
M3 - Journal article
C2 - 22882974
VL - 13
SP - 1532
EP - 1546
JO - Traffic
JF - Traffic
SN - 1398-9219
IS - 11
ER -
ID: 44381516