Insulin-induced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor in adipocytes in vitro

Takanobu Yamamoto, Takayoshi Suganami, Minako Kiso-Narita, Peggy A. Scherle, Yasutomi Kamei, Mitsuaki Isobe, Shigeki Higashiyama, Yoshihiro Ogawa

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is synthesized as a type I transmembrane protein, which is proteolytically cleaved to release a soluble form via members of the a disintegrin and metalloproteinase (ADAM) family of proteolytic enzymes. This study was designed to elucidate the molecular mechanism underlying insulin-induced HB-EGF shedding in adipocytes in vitro. The 3T3-L1 adipocytes with stable expression of alkaline phosphatase (AP)-tagged proHB-EGF (3T3-L1/HB-EGF-AP adipocytes) were developed and AP activities of conditioned media were determined. Using 3T3-L1/HB-EGF-AP adipocytes, we demonstrated that insulin induces HB-EGF shedding in differentiated 3T3-L1 adipocytes in a dose- and time-dependent manner. There is no significant increase in insulin-induced HB-EGF shedding in undifferentiated 3T3-L1 preadipocytes. Studies with metalloprotease inhibitors suggested that insulin-induced HB-EGF shedding in adipocytes is mediated at least in part via ADAM17. Treatment with recombinant HB-EGF results in a dose- and time-dependent increase in HB-EGF shedding in adipocytes, which is significantly suppressed by pharmacologic blockade of ADAM17 (P < 0.01). Moreover, insulin-induced HB-EGF shedding in adipocytes is significantly inhibited by AG1478, an EGF receptor antagonist (P < 0.01). This study provides in vitro evidence that insulin induces HB-EGF shedding in 3T3-L1 adipocytes. Our data also suggest the role of ADAM17 in insulin-induced HB-EGF shedding in adipocytes.

Original languageEnglish
Pages (from-to)1888-1894
Number of pages7
JournalObesity
Volume18
Issue number10
DOIs
Publication statusPublished - Oct 1 2010
Externally publishedYes

Fingerprint

Epidermal Growth Factor
Adipocytes
Heparin
Intercellular Signaling Peptides and Proteins
Insulin
Alkaline Phosphatase
Metalloproteases
In Vitro Techniques
Disintegrins
Conditioned Culture Medium
Epidermal Growth Factor Receptor
Peptide Hydrolases

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Nutrition and Dietetics

Cite this

Insulin-induced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor in adipocytes in vitro. / Yamamoto, Takanobu; Suganami, Takayoshi; Kiso-Narita, Minako; Scherle, Peggy A.; Kamei, Yasutomi; Isobe, Mitsuaki; Higashiyama, Shigeki; Ogawa, Yoshihiro.

In: Obesity, Vol. 18, No. 10, 01.10.2010, p. 1888-1894.

Research output: Contribution to journalArticle

Yamamoto, T, Suganami, T, Kiso-Narita, M, Scherle, PA, Kamei, Y, Isobe, M, Higashiyama, S & Ogawa, Y 2010, 'Insulin-induced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor in adipocytes in vitro', Obesity, vol. 18, no. 10, pp. 1888-1894. https://doi.org/10.1038/oby.2010.2
Yamamoto T, Suganami T, Kiso-Narita M, Scherle PA, Kamei Y, Isobe M et al. Insulin-induced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor in adipocytes in vitro. Obesity. 2010 Oct 1;18(10):1888-1894. https://doi.org/10.1038/oby.2010.2
Yamamoto, Takanobu ; Suganami, Takayoshi ; Kiso-Narita, Minako ; Scherle, Peggy A. ; Kamei, Yasutomi ; Isobe, Mitsuaki ; Higashiyama, Shigeki ; Ogawa, Yoshihiro. / Insulin-induced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor in adipocytes in vitro. In: Obesity. 2010 ; Vol. 18, No. 10. pp. 1888-1894.
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