The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme.

The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme. Research Abstract Details 

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The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme. Abstract Text:

Abstract The N and C domains of somatic angiotensin-converting enzyme (sACE) differ in terms of their substrate specificity, inhibitor profiling, chloride dependency and thermal stability. The C domain is thermally less stable than sACE or the N domain. Since both domains are heavily glycosylated, the effect of glycosylation on their thermal stability was investigated by assessing their catalytic and physicochemical properties. Testis ACE (tACE) expressed in mammalian cells, mammalian cells in the presence of a glucosidase inhibitor and insect cells yielded proteins with altered catalytic and physicochemical properties, indicating that the more complex glycans confer greater thermal stabilization. Furthermore, a decrease in tACE and N-domain N-glycans using site-directed mutagenesis decreased their thermal stability, suggesting that certain N-glycans have an important effect on the protein's thermodynamic properties. Evaluation of the thermal stability of sACE domain swopover and domain duplication mutants, together with sACE expressed in insect cells, showed that the C domain contained in sACE is less dependent on glycosylation for thermal stabilization than a single C domain, indicating that stabilizing interactions between the two domains contribute to the thermal stability of sACE and are decreased in a C-domain-duplicating mutant.

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The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Biological chemistry

VOLUME: 389

Page Numbers: 1153-61

Journal Abbreviation: Biol. Chem.

ISSN: 1431-6730

DAY: 18

MONTH: Sep

YEAR: 2008

The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme. Information

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LANGUAGE: eng

NlmUniqueID: 9700112

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Grant and Affiliation Information for The role of glycosylation and domain interactions in the thermal stability of human angiotensin-converting enzyme.

AFFILIATION: 1Division of Medical Biochemistry, Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Observatory 7925, South Africa.

Country: Germany

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MEDLINETA: Biol Chem

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