Unit:
Τομέας Βασικών ΕπιστημώνLibrary of the School of Health Sciences
Author:
Βασιλάκος Γεώργιος
Dissertation committee:
Αν. Καθ. Ε. Κοτσιφάκη
Original Title:
'Εκφραση του πεπτιδίου επέκτασης (Ε-peptide) του IGF-1 στην κυτταρική σειρά SK-UT-1 και σε ιστούς ανθρώπου και τρωκτικών
Summary:
Insulin-like growth factor-1 (IGF-1) has endocrine and autocrine/paracrine
actions that regulates the pre- and postnatal development in many cell types
and tissues of both human and rodent origin. The igf-1 gene, contains six
exons, and gives rise to multiple, heterogeneous transcripts (mRNA) by a
combination of: a) alternative usage of promoters, b) alternative splicing and
c) different polyadenylation signals. These multiple IGF-1 transcripts encode
different precursor peptides, which could undergo post-translational
modification. Briefly, are encoded the mature IGF-1 peptide and the E-peptides.
In humans are potentially produced three E-peptides (Ea, Eb and Ec), while in
rodents two (Ea and Eb). Therefore, the objectives of the present study are to
demonstrate the existence of Ec/Eb peptides and investigate their biological
activity in an in vitro model. By immunobloting, with a polyclonal anti-IGF-1Ec
antibody, we detected the pro-form and the E-peptide in various tissues and
verified that the peptide can be derived from proteolysis of the precursor
peptide, through the action of the furin enzyme. To examine the bioactivity of
the peptide, a synthetic Eb and Scramble peptide were used. Additional by
developing a ligand binding in vitro assay we investigated, whether the
synthetic Eb peptide Eb can bind receptor. Finally, we describe a cellular
model (SK-UT-1) which does not endogenously express IGF-1 and IGF-1R, and could
be utilized for elucidating the biological role of the peptides, independently
of the mature IGF-1 peptide.
Keywords:
IGF-1, Ε-peptide, Ec/Eb, SK-UT-1, Alternative splicing
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