Unit:
Τομέας Βιοχημείας Μοριακής ΒιολογίαςLibrary of the School of Science
Author:
Ζερβακάκη Βασιλική
Supervisors info:
Βασιλακοπούλου Διδώ Αναπλ. Καθηγ. (Επιβλέπουσα), Σίδερης Διαμάντης Αναπλ. Καθηγ., Ευθυμιόπουλος Σπύρος Αναπλ. Καθηγ.
Original Title:
Μελέτη της κυτταρικής τοπολογίας της πλήρους μήκους και των εναλλακτικών ισομορφών της L-Dopa αποκαρβοξυλάσης
Translated title:
Study upon thw subcellylar topology of full-length l- dopa dycarboxylase and alternative isoforms
Summary:
The goals of this Project were: 1) Investigation of the cellular topology of
full-length DDC and alternative isoforms alt-DDC and DDC lacking exon 3 [DDC
(-3)], using as cell model the cell line CHO which does not express
endogenously DDC, 2) Investigation of the release mechanisms and the and
influence of different factors on the solubilization of DDC and DDC (-3). DDC
and DDC (-3) protein molecules were detected in all three separation phases.
The alternative isoform alt-DDC was not recovered in the hydrophovic phase. DDC
was released in the soluble supernatant after incubation with pH 4. On the
other hand, DDC (-3) was released after incubation with pH 6. Both isoforms
released after incubation time between 30 and 60 minutes. Our data indicated
that only a portion of the membrane-associated DDC molecules were released into
the soluble fraction.It was observed that increasing concentrations of
aprotinin cause an inhibitory effect on both isoforms. Ιncreasing
concentrations of divalent cations, were different in both cell lines,
suggesting the complexity in regulation of two molecules. It is noteworthy that
in the DDC (-3) experiments, were observed phenomena of possible
post-translational modification. In conclusion, it is necessary to further
investigate the above phenomena to better understand the complex mechanisms
regulating the enzyme DDC and alternative isoforms.
Keywords:
L-DOPA decarboxylase, Topology of DDC, Topology of DDC alternative isoforms, Release mechanisms of DDC, Triton x-114
Number of references:
226
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