KOPS - Das Institutionelle Repositorium der Universität Konstanz

Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli

Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli

Zitieren

Dateien zu dieser Ressource

Prüfsumme: MD5:44ef062cd760669dd04635982c31b783

BROSS, Peter, Stefan ENGST, Arnold W. STRAUSS, Daniel P. KELLY, Ihab RASCHED, Sandro GHISLA, 1990. Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli. In: The Journal of Biological Chemistry. 265(13), pp. 7116-7119

@article{Bross1990Chara-7533, title={Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli}, year={1990}, number={13}, volume={265}, journal={The Journal of Biological Chemistry}, pages={7116--7119}, author={Bross, Peter and Engst, Stefan and Strauss, Arnold W. and Kelly, Daniel P. and Rasched, Ihab and Ghisla, Sandro} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/7533"> <dc:creator>Strauss, Arnold W.</dc:creator> <dc:contributor>Engst, Stefan</dc:contributor> <dc:contributor>Bross, Peter</dc:contributor> <dc:contributor>Ghisla, Sandro</dc:contributor> <dc:contributor>Rasched, Ihab</dc:contributor> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:35:11Z</dc:date> <dcterms:rights rdf:resource="https://creativecommons.org/licenses/by-nc-nd/2.0/legalcode"/> <dcterms:title>Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli</dcterms:title> <dc:language>eng</dc:language> <dc:creator>Kelly, Daniel P.</dc:creator> <dcterms:abstract xml:lang="eng">The cDNA of human medium chain acyl-CoA dehydrogenase (MCADH) was modified by in vitro mutagenesis, and the sequence encoding the mature form of MCADH was introduced into an inducible expression plasmid. We observed synthesis of the protein in Escherichia coli cells transformed with this plasmid with measurable MCADH enzyme activity in cell extracts. Glutamic acid 376, which has been proposed by Powell and Thorpe (Powell, P. J., and Thorpe, J. (1988) Biochemistry 27, 8022- 8028) as an essential residue and the proton-abstracting base at the active site of the enzyme, was mutated to glutamine. After expression in bacteria of this plasmid, the corresponding extracts show no detectable MCADH activity, although mutant MCADH-protein production was detected by protein immunoblots. The mature enzyme and the Gln376 mutant were purified to apparent homogeneity. The wild-type enzyme is a yellow protein due to the content of stoichiometric FAD and had a specific activity which is 50% of MCADH purified from pig kidney. The Gln376 mutant is devoid of activity (less than 0.02% that of wild type, expressed enzyme) and is green because of bound CoA persulfide. Properties of the mutant enzyme suggest that the Glu376→Gln change specifically affects substrate binding. These results prove that Glu376 plays an important role in the initial step of dehydrogenation catalysis.</dcterms:abstract> <dc:creator>Ghisla, Sandro</dc:creator> <dc:creator>Rasched, Ihab</dc:creator> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:35:11Z</dcterms:available> <dc:creator>Bross, Peter</dc:creator> <dcterms:bibliographicCitation>First publ. in: The Journal of Biological Chemistry 265 (1990), 13, pp. 7116-7119</dcterms:bibliographicCitation> <dc:creator>Engst, Stefan</dc:creator> <dc:contributor>Strauss, Arnold W.</dc:contributor> <dc:format>application/pdf</dc:format> <dcterms:issued>1990</dcterms:issued> <dc:rights>deposit-license</dc:rights> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/7533"/> <dc:contributor>Kelly, Daniel P.</dc:contributor> </rdf:Description> </rdf:RDF>

Dateiabrufe seit 01.10.2014 (Informationen über die Zugriffsstatistik)

J_Biol_Chem_1990_BrossCharacterization_of_wild_type_and_an.pdf 108

Das Dokument erscheint in:

deposit-license Solange nicht anders angezeigt, wird die Lizenz wie folgt beschrieben: deposit-license

KOPS Suche


Stöbern

Mein Benutzerkonto