Linked Life Data

18297792

Source:http://linkedlifedata.com/resource/pubmed/id/18297792

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2008-2-26
pubmed:abstractText
We improved an already existing cytochrome c expression system to a reliable, tightly controllable one to achieve a higher expression yield for single cysteine mutants of horse cytochrome c. The protein is heterologously overexpressed in E. coli together with the maturation coordinating enzyme heme lyase from yeast. Various plasmid constructs and host strains were tested for protein expression yield and routinely around 35 mg/L yield was achieved, which is a good result for a post-translationally modified enzyme. The purpose of producing cysteine mutants is to position accessible cysteine residues on the surface of cytochrome c which can be labeled with a photoactive redox dye, 8-thiouredopyrene-1,3,6-trisulfonate, TUPS. TUPS labeled proteins have been used for intramolecular and intermolecular electron transfer measurements. Here, we initiate the photoreduction of cytochrome c oxidase, the natural electron acceptor partner of cytochrome c by an appropriate cytochrome c mutant labeled with TUPS. The electron transfer from cytochrome c to the first cytochrome oxidase redox cofactor, copper A, is shown to be very fast.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
0236-5383
pubmed:author
pubmed:issnType
Print
pubmed:volume
58 Suppl
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
23-35
pubmed:meshHeading
pubmed:year
2007
pubmed:articleTitle
Improved system for heterologous expression of cytochrome c mutants in Escherichia coli.
pubmed:affiliation
Institute of Biophysics, Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't