Identification of the cleavage sites of transforming growth factor α by insulin-degrading enzymes

Frederick G Hamel, Barry D. Gehm, Marsha Rich Rosner, William C. Duckworth

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Insulin-degrading enzyme (IDE) is a sulfhydryl-dependent metalloproteinase with a zinc binding site unique to a new class of proteinases. The enzyme is relatively specific for a number of hormones/growth factors, such as insulin, atrial natriuretic peptide, IGF-II, and proinsulin. In this study we have identified the amino-acid bonds cleaved by IDE in transforming growth factor-α. High-performance liquid chromatography was used to separate the peptides generated by the degradation of 125I-TGF-α. The peptides were then submitted to sequential Edman degradation to determine the peptide bond broken. Cleavage sites were found at amino acids, 10-11 (Asp-Ser), 25-26 (Val-Gln), 28-29 (Asp-Lys), and 30-31 (Pro-Ala). In agreement with studies of cleavage sites of other hormones by this enzyme, no clear amino-acid specificity was seen. However, examination of the sites on a three-dimensional model of TGF-α suggest the primary mechanism used by IDE for determining cleavage sites is the tertiary structure of the substrate.

Original languageEnglish (US)
Pages (from-to)207-214
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1338
Issue number2
DOIs
StatePublished - Apr 4 1997

Fingerprint

Insulysin
Transforming Growth Factors
aspartyllysine
Amino Acids
Peptides
Hormones
Proinsulin
Degradation
Insulin-Like Growth Factor II
High performance liquid chromatography
Metalloproteases
Atrial Natriuretic Factor
Enzymes
Zinc
Intercellular Signaling Peptides and Proteins
Peptide Hydrolases
Binding Sites
High Pressure Liquid Chromatography
Insulin
Substrates

Keywords

  • Insulin-degrading enzyme
  • Insulysin
  • Metalloproteinase
  • Sulfhydryl
  • Transforming growth factor α
  • Zinc binding site

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

Cite this

Identification of the cleavage sites of transforming growth factor α by insulin-degrading enzymes. / Hamel, Frederick G; Gehm, Barry D.; Rosner, Marsha Rich; Duckworth, William C.

In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, Vol. 1338, No. 2, 04.04.1997, p. 207-214.

Research output: Contribution to journalArticle

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