Chapter title |
Identification of Proteolytic Cleavage Sites of EphA2 by Membrane Type 1 Matrix Metalloproteinase on the Surface of Cancer Cells
|
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Chapter number | 3 |
Book title |
Proteases and Cancer
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7595-2_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7594-5, 978-1-4939-7595-2
|
Authors |
Keiji Kikuchi, Hiroko Kozuka-Hata, Masaaki Oyama, Motoharu Seiki, Naohiko Koshikawa |
Abstract |
Proteolytic cleavage of membrane proteins can alter their functions depending on the cleavage sites. We recently demonstrated that membrane type 1 matrix metalloproteinase (MT1-MMP ) converts the tumor suppressor EphA2 into an oncogenic signal transducer through EphA2 cleavage. The cleaved EphA2 fragment that remains at the cell surface may be a better target for cancer therapy than intact EphA2. To analyze the cleavage site(s) of EphA2, we purified the fragments from tumor cells expressing MT1-MMP and Myc- and 6× His-tagged EphA2 by two-step affinity purification . The purified fragment was digested with trypsin to generate proteolytic peptides , and the amino acid sequences of these peptides were determined by nano-LC-mass spectrometry to identify the MT1-MMP-mediated cleavage site(s) of EphA2. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 4 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 1 | 25% |
Unknown | 3 | 75% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 1 | 25% |
Medicine and Dentistry | 1 | 25% |
Unknown | 2 | 50% |