Electrochemical detection of receptor-mediated endocytosis by scanning electrochemical microscopy

Yasufumi Takahashi; Takeshi Miyamoto; Hitoshi Shiku; Kosuke Ino; Tomoyuki Yasukawa; Ryutaro Asano; Izumi Kumagai; Tomokazu Matsue

doi:10.1039/c1cp21886g

Electrochemical detection of receptor-mediated endocytosis by scanning electrochemical microscopy

Yasufumi Takahashi, Takeshi Miyamoto, Hitoshi Shiku, Kosuke Ino, Tomoyuki Yasukawa, Ryutaro Asano, Izumi Kumagai, Tomokazu Matsue

Research output: Contribution to journal › Article › peer-review

17 Citations (Scopus)

Abstract

We report a scanning electrochemical microscopy (SECM)-based receptor-mediated endocytosis detection method. Epidermal growth factor receptor (EGFR), which is one of the key membrane proteins associated with cancer, was used as a model for receptor-mediated endocytosis. EGFR molecules on the outer cell membrane were detected by SECM by using alkaline phosphatase (ALP) as a labeling enzyme. Since SECM detected the ALP activity on the outer membrane, the procedure helped discriminate the EGFR on the outer membrane from the intracellular EGFR involved in endocytosis. SECM showed a marked decrease in the current responses generated due to ALP activity by 93% on addition of the epidermal growth factor, indicating clearly that EGF triggered the endocytosis, which led to the withdrawal of most EGFRs from the outer membrane.

Original language	English
Pages (from-to)	16569-16573
Number of pages	5
Journal	Physical Chemistry Chemical Physics
Volume	13
Issue number	37
DOIs	https://doi.org/10.1039/c1cp21886g
Publication status	Published - 2011 Oct 7

ASJC Scopus subject areas

Physics and Astronomy(all)
Physical and Theoretical Chemistry

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10.1039/c1cp21886g

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abstract = "We report a scanning electrochemical microscopy (SECM)-based receptor-mediated endocytosis detection method. Epidermal growth factor receptor (EGFR), which is one of the key membrane proteins associated with cancer, was used as a model for receptor-mediated endocytosis. EGFR molecules on the outer cell membrane were detected by SECM by using alkaline phosphatase (ALP) as a labeling enzyme. Since SECM detected the ALP activity on the outer membrane, the procedure helped discriminate the EGFR on the outer membrane from the intracellular EGFR involved in endocytosis. SECM showed a marked decrease in the current responses generated due to ALP activity by 93% on addition of the epidermal growth factor, indicating clearly that EGF triggered the endocytosis, which led to the withdrawal of most EGFRs from the outer membrane.",

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AU - Takahashi, Yasufumi

AU - Miyamoto, Takeshi

AU - Shiku, Hitoshi

AU - Ino, Kosuke

AU - Yasukawa, Tomoyuki

AU - Asano, Ryutaro

AU - Kumagai, Izumi

AU - Matsue, Tomokazu

PY - 2011/10/7

Y1 - 2011/10/7

N2 - We report a scanning electrochemical microscopy (SECM)-based receptor-mediated endocytosis detection method. Epidermal growth factor receptor (EGFR), which is one of the key membrane proteins associated with cancer, was used as a model for receptor-mediated endocytosis. EGFR molecules on the outer cell membrane were detected by SECM by using alkaline phosphatase (ALP) as a labeling enzyme. Since SECM detected the ALP activity on the outer membrane, the procedure helped discriminate the EGFR on the outer membrane from the intracellular EGFR involved in endocytosis. SECM showed a marked decrease in the current responses generated due to ALP activity by 93% on addition of the epidermal growth factor, indicating clearly that EGF triggered the endocytosis, which led to the withdrawal of most EGFRs from the outer membrane.

AB - We report a scanning electrochemical microscopy (SECM)-based receptor-mediated endocytosis detection method. Epidermal growth factor receptor (EGFR), which is one of the key membrane proteins associated with cancer, was used as a model for receptor-mediated endocytosis. EGFR molecules on the outer cell membrane were detected by SECM by using alkaline phosphatase (ALP) as a labeling enzyme. Since SECM detected the ALP activity on the outer membrane, the procedure helped discriminate the EGFR on the outer membrane from the intracellular EGFR involved in endocytosis. SECM showed a marked decrease in the current responses generated due to ALP activity by 93% on addition of the epidermal growth factor, indicating clearly that EGF triggered the endocytosis, which led to the withdrawal of most EGFRs from the outer membrane.

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Electrochemical detection of receptor-mediated endocytosis by scanning electrochemical microscopy

Abstract

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