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Biochemical characterization of endothelin-converting enzyme-1alpha in cultured skin-derived cells and its postulated

Akira Hachiya¹, Takeshi Kobayashi, Yoshinori Takema

¹Kao Biological Science Laboratories, Ichikaimachi 2606, Tochigi 321-3497, Japan.

The Journal of Biological Chemistry

|November 28, 2001

Summary

Endothelin-converting enzyme-1alpha (ECE-1alpha) processes and secretes endothelin-1 (ET-1) in human skin keratinocytes. This enzyme plays a key role in UVB-induced pigmentation and may link to post-inflammatory hyperpigmentation.

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Area of Science:

Dermatology
Biochemistry
Molecular Biology

Background:

Endothelin-1 (ET-1) stimulates skin pigmentation, with increased secretion by keratinocytes after UVB exposure.
The biological mechanisms of ET-1 secretion by keratinocytes remain unclear.
Endothelin-converting enzyme (ECE) processes inactive Big ET to active ET in endothelial cells.

Purpose of the Study:

To investigate the role of ECE in the processing and secretion of ET-1 by human keratinocytes.
To characterize the expression and activity of ECE-1alpha in human skin cells.

Main Methods:

Synthesized a peptide of human ECE-1alpha and generated a specific antibody (alphaPEPT6).
Utilized reverse transcription PCR and Western blotting to detect ECE-1 transcripts and protein.
Assayed ECE activity in keratinocyte extracts across various pHs and tested the effect of alphaPEPT6.

Main Results:

Human keratinocytes express significant levels of ECE-1 transcripts and ECE-1alpha protein.
ECE activity in keratinocytes showed an optimal neutral pH of 7.0 and was pH-dependent.
The antibody alphaPEPT6 significantly reduced ECE activity, confirming ECE-1alpha's role.
Interleukin-1alpha increased ECE-1 transcript expression, suggesting a link to post-inflammatory hyperpigmentation.

Conclusions:

ECE-1alpha is expressed in human skin cells, including keratinocytes.
ECE-1alpha is crucial for the processing and UVB-inducible secretion of ET-1.
These findings highlight ECE-1alpha's role in epidermal pigmentation and potential involvement in hyperpigmentation disorders.