Mint-3 regulates the retrieval of the internalized membrane-type matrix metalloproteinase, MT5-MMP, to the plasma membrane by binding to its carboxyl end motif EWV.
Membrane type matrix metalloproteinases (MT-MMPs) play a critical role in promoting cell growth and migration within the extracellular matrix by trafficking to specialized areas. Here we show that the carboxyl EWV motif of MT5-MMP serves as a retrieval signal for internalized MT5-MMP by interacting with Mint-3, a protein with two type III PDZ domains. Deletion of the EWV signal impairs the recycling of MT5-MMP without affecting its internalization, leading to decreased activity on the cell surface. A yeast two-hybrid screening identified Mint-3 as the EWV-binding protein. Mint-3 stimulates MT5-MMP activity when expressed at low levels in an EWV-dependent fashion, but inhibits its activity at higher levels independent of the EWV motif. siRNA-mediated knockdown of endogenous Mint-3 decreased MT5-MMP activity. Furthermore, Mint-3 significantly increased the level of MT5-MMP on the cell surface without affecting its synthesis and internalization. Therefore, Mints may be the adaptor proteins that regulate the trafficking of MT-MMPs.
Department of Pharmacology, University of Minnesota School of Medicine, Minneapolis, MN 55455, USA.
Ping
P
Wang
Xing
X
Wang
Duanqing
D
Pei
D048868:Adaptor Proteins, Signal Transducing; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001710:Biotin / Q000378:metabolism; D015153:Blotting, Western; D002352:Carrier Proteins; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000201:enzymology* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D034281:Dynamins / Q000737:chemistry; D017353:Gene Deletion; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000378:metabolism*; D018613:Microscopy, Confocal; D008954:Models, Biological; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism / Q000502:physiology*; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D020798:Two-Hybrid System Techniques; D021601:trans-Golgi Network / Q000378:metabolism
C117536:APBA3 protein, human; D048868:Adaptor Proteins, Signal Transducing; C485530:Apba3 protein, mouse; D002352:Carrier Proteins; D011506:Proteins; D034741:RNA, Small Interfering; D001710:Biotin; D005982:Glutathione Transferase; C492243:MMP24 protein, human; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D034281:Dynamins