Interaction of the Metalloprotease Disintegrins MDC9 and MDC15 with Two SH3 Domain-containing Proteins, Endophilin I and SH3PX1*
- From the ‡Cellular Biochemistry and Biophysics Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York 10021 and ‖Respiratory and Inflammation Research, AstraZeueca Pharmaceuticals, Alderley Park, Macclesfield, Cheshire SK 10 4TG, United Kingdom
Abstract
Metalloprotease disintegrins (a disintegrin and metalloprotease (ADAM) and metalloprotease, disintegrin,cysteine-rich proteins (MDC)) are a family of membrane-anchored glycoproteins that function in diverse biological processes, including fertilization, neurogenesis, myogenesis, and ectodomain processing of cytokines and other proteins. The cytoplasmic domains of ADAMs often include putative signaling motifs, such as proline-rich SH3 ligand domains, suggesting that interactions with cytoplasmic proteins may affect metalloprotease disintegrin function. Here we report that two SH3 domain-containing proteins, endophilin I (SH3GL2, SH3p4) and a novel SH3 domain- andphox homology (PX) domain-containing protein, termed SH3PX1, can interact with the cytoplasmic domains of the metalloprotease disintegrins MDC9 and MDC15. These interactions were initially identified in a yeast two-hybrid screen and then confirmed using bacterial fusion proteins and co-immunoprecipitations from eukaryotic cells expressing both binding partners. SH3PX1 and endophilin I both preferentially bind the precursor but not the processed form of MDC9 and MDC15 in COS-7 cells. Since rat endophilin I is thought to play a role in synaptic vesicle endocytosis and SH3PX1 has sequence similarity to sorting nexins in yeast, we propose that endophilin I and SH3PX1 may have a role in regulating the function of MDC9 and MDC15 by influencing their intracellular processing, transport, or final subcellular localization.
Footnotes
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↵* This work was supported in part by National Institutes of Health Grant R55GM51988 (to C. P. B. for MDC9), by National Research Service Award 5F32GM18585-02 (to K. K. N.), by the DeWitt Wallace Fund for Memorial Sloan-Kettering Cancer Center (MSKCC), and by MSKCC Grant NCI-P30-CA-08748.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) , and .
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↵§ Both authors contributed equally to this work.
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↵¶ Supported by the Zeneca Strategic Research Fund (for MDC15 as part of a grant to C. P. B.).
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↵** To whom correspondence should be addressed: Cellular Biochemistry and Biophysics Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, Box 368, 1275 York Ave., New York, NY 10021. Tel.: 212-639-2915; Fax: 212-717-3047; E-mail: c-blobel@ski.mskcc.org.
- Abbreviations:
- ADAMs
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a disintegrin and metalloprotease
- MDC
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metalloprotease, disintegrin,cysteine-rich proteins
- PCR
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polymerase chain reaction
- GST
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glutathione S-transferase
- TACE
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tumor necrosis factor α-converting enzyme
- HB-EGF
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heparin-binding epidermal growth factor-like growth factor
- SH3
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Src homology 3
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- Received May 7, 1999.
- Revision received July 23, 1999.
- The American Society for Biochemistry and Molecular Biology, Inc.
