Proteoglykane und die Verpackung von Exportproteinen: Interaction von Serglycin und ZG 16 in den Zymogengranula des exokrinen Rattenpankreas
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Philipps-Universität Marburg
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Abstract
In this thesis the interaction of the mainly
membrane-associated secretory lectin ZG 16 and the proteoglycan
serglycin should be analysed to gain deeper insight in the
mechanisms of sorting and packaging of the zymogens into
secretory granules. Moreover further proteoglycans should be
isolated and identified from the content fraction of the
zymogen granules.
The following results were obtained:
1.
By cloning and expressing the unmodified N- and C-terminal
parts of serglycin it could be demonstrated in various
protein-protein-interaction studies that serglycin interacts
with ZG 16 via its unmodified N-terminal part (SGN) and is so
linked to the granule membrane.
2. By analysing several
point- and deletion-mutants of the N-terminal part of serglycin
(SGN), the binding sequence could be restricted to 9 amino
acids (ARYQWVRCK) near the N-terminus of serglycin.
3.
Modelling the hypothetical secondary structure of serglycin
(ExPASy Molecular Biology Server) it was revealed that the
binding motif of serglycin to ZG 16 forms a b-strand.
This
interaction represents a sorting mechanism by which the
zymogens, that are attached to the glycosaminoglycans of
serglycin (as previously shown by Biederbick et al., 2003) are
transported into the zymogen granules.
4. Further
proteoglycans in the granule content could only be shown
indirectly. In a special proteoglycan precipitation assay
(Blyscan Assay) proteoglycans could be detected by extinction
measurement. The highest extinction was obtained after
incubating the content fraction with NaHCO3.In this thesis the interaction of the mainly
membrane-associated secretory lectin ZG 16 and the proteoglycan
serglycin should be analysed to gain deeper insight in the
mechanisms of sorting and packaging of the zymogens into
secretory granules. Moreover further proteoglycans should be
isolated and identified from the content fraction of the
zymogen granules.
The following results were obtained:
1.
By cloning and expressing the unmodified N- and C-terminal
parts of serglycin it could be demonstrated in various
protein-protein-interaction studies that serglycin interacts
with ZG 16 via its unmodified N-terminal part (SGN) and is so
linked to the granule membrane.
2. By analysing several
point- and deletion-mutants of the N-terminal part of serglycin
(SGN), the binding sequence could be restricted to 9 amino
acids (ARYQWVRCK) near the N-terminus of serglycin.
3.
Modelling the hypothetical secondary structure of serglycin
(ExPASy Molecular Biology Server) it was revealed that the
binding motif of serglycin to ZG 16 forms a b-strand.
This
interaction represents a sorting mechanism by which the
zymogens, that are attached to the glycosaminoglycans of
serglycin (as previously shown by Biederbick et al., 2003) are
transported into the zymogen granules.
4. Further
proteoglycans in the granule content could only be shown
indirectly. In a special proteoglycan precipitation assay
(Blyscan Assay) proteoglycans could be detected by extinction
measurement. The highest extinction was obtained after
incubating the content fraction with NaHCO3.
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Dates
Created: 2003Issued: 2003-12-22Updated: 2021-07-14
Faculty
Medizin
Publisher
Philipps-Universität Marburg
Language
ger
Data types
DoctoralThesis
Keywords
ProteoglycansZymogeneSerglycinSerglycinZymogenesProteinsortierungProtein Sorting
DFG-subjects
Granulum GlykosaminoglykaneProteoglykane
DDC-Numbers
610
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Hillebrand, Merle (128603178): Proteoglykane und die Verpackung von Exportproteinen: Interaction von Serglycin und ZG 16 in den Zymogengranula des exokrinen Rattenpankreas. : Philipps-Universität Marburg 2003-12-22. DOI: https://doi.org/10.17192/z2003.0709.
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This item has been published with the following license: In Copyright