TY - JOUR
T1 - Identification of the cleavage sites of transforming growth factor α by insulin-degrading enzymes
AU - Hamel, Frederick G.
AU - Gehm, Barry D.
AU - Rosner, Marsha Rich
AU - Duckworth, William C.
N1 - Funding Information:
The authors thank Mark Hermondson of Purdue University and the University of Nebraska Protein Sequencing Facility for peptide sequencing. The authors would also like to thank Carol Peters for her assistance in preparing this manuscript. This work was supported in part by the Department of Veterans Affairs Research Service.
PY - 1997/4/4
Y1 - 1997/4/4
N2 - 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.
AB - 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.
KW - Insulin-degrading enzyme
KW - Insulysin
KW - Metalloproteinase
KW - Sulfhydryl
KW - Transforming growth factor α
KW - Zinc binding site
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U2 - 10.1016/S0167-4838(96)00202-6
DO - 10.1016/S0167-4838(96)00202-6
M3 - Article
C2 - 9128138
AN - SCOPUS:0030951993
SN - 0167-4838
VL - 1338
SP - 207
EP - 214
JO - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
JF - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
IS - 2
ER -