Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin
| ISSN: | 0264-6021 |
|---|---|
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f92651f3266723f24cbcd05fd259a71b https://pubmed.ncbi.nlm.nih.gov/7848297 |
| Rights: | OPEN |
| Přírůstkové číslo: | edsair.doi.dedup.....f92651f3266723f24cbcd05fd259a71b |
| Autor: | J F Briat, O Van Wuytswinkel |
| Rok vydání: | 1995 |
| Předmět: |
Protein Denaturation
Protein Conformation Protein subunit Mutant Molecular Sequence Data Biology Ferroxidase activity Biochemistry Protein Structure Secondary law.invention Structure-Activity Relationship law Urea Denaturation (biochemistry) Amino Acid Sequence Site-directed mutagenesis Molecular Biology Plant Proteins chemistry.chemical_classification Base Sequence Peas food and beverages Cell Biology Molecular biology Peptide Fragments Recombinant Proteins Amino acid Ferritin chemistry Ferritins Seeds biology.protein Recombinant DNA Mutagenesis Site-Directed Thermodynamics Research Article |
| Zdroj: | The Biochemical journal. 305 |
| ISSN: | 0264-6021 |
| Popis: | Plant ferritin has a three-dimensional structure predicted to be very similar to that of animal ferritin. It has, however, an additional specific sequence of 24 amino acids at its N-terminus named extension peptide (EP). In order to determine precisely the interactions between EP and other domains of the pea seed ferritin subunit, three point mutations were performed. The mutated residues were chosen by three-dimensional computer modelling of the pea seed ferritin subunit structure [Lobréaux, Yewdall, Briat and Harrison (1992) Biochem. J. 228, 931-939]. The mutant recombinant proteins were expressed in Escherichia coli and purified to homogeneity; all the mutants were found to be assembled as 24-mers. When Ala-13 was replaced by His, as in mammalian ferritins, ferroxidase activity was significantly reduced. Moreover, in vitro iron-core formation in Pro-X-->Ala, Lys-R-->Glu and Ala-13-->His mutants was increased after denaturation by urea followed by renaturation; this was also observed with the EP deletion mutant (r delta TP/EP). The recombinant ferritins were also analysed using tryptophan fluorescence spectra. The r delta TP/EP, Pro-X-->Ala and Lys-R-->Glu mutants were found to be more susceptible to denaturation by urea than the native r delta TP pea seed ferritin. |
| Databáze: | OpenAIRE |
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