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Wyszukujesz frazę "site directed mutagenesis" wg kryterium: Temat


Wyświetlanie 1-2 z 2
Tytuł:
In vitro fluorescence studies of transcription factor IIB-DNA interaction
Autorzy:
Górecki, Andrzej
Figiel, Małgorzata
Dziedzicka-Wasylewska, Marta
Powiązania:
https://bibliotekanauki.pl/articles/1038975.pdf
Data publikacji:
2015
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
transcription factors
TF2B
TFIIB
fluorescence spectroscopy
fluorescence quenching
site directed mutagenesis
Opis:
General transcription factor TFIIB is one of the basal constituents of the preinitiation complex of eukaryotic RNA polymerase II, acting as a bridge between the preinitiation complex and the polymerase, and binding promoter DNA in an asymmetric manner, thereby defining the direction of the transcription. Methods of fluorescence spectroscopy together with circular dichroism spectroscopy were used to observe conformational changes in the structure of recombinant human TFIIB after binding to specific DNA sequence. To facilitate the exploration of the structural changes, several site-directed mutations have been introduced altering the fluorescence properties of the protein. Our observations showed that binding of specific DNA sequences changed the protein structure and dynamics, and TFIIB may exist in two conformational states, which can be described by a different microenvironment of W52. Fluorescence studies using both intrinsic and exogenous fluorophores showed that these changes significantly depended on the recognition sequence and concerned various regions of the protein, including those interacting with other transcription factors and RNA polymerase II. DNA binding can cause rearrangements in regions of proteins interacting with the polymerase in a manner dependent on the recognized sequences, and therefore, influence the gene expression.
Źródło:
Acta Biochimica Polonica; 2015, 62, 3; 413-421
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Probing iso-1-cytochrome c structure by site-directed spin labeling and electron paramagnetic resonance techniques
Autorzy:
Pyka, Janusz
Osyczka, Artur
Turyna, Bohdan
Blicharski, Wojciech
Froncisz, Wojciech
Powiązania:
https://bibliotekanauki.pl/articles/1044446.pdf
Data publikacji:
1999
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
iso-1-cytochrome c
Saccharomyces cerevisiae
site-directed mutagenesis
spin label
electron paramagnetic resonance
Opis:
A cysteine-specific methanethiosulfonate spin label was introduced into yeast iso-1-cytochrome c at three different positions. The modified forms of cytochrome c included: the wild-type protein labeled at naturally occurring C102, and two mutated proteins, S47C and L85C, labeled at positions 47 and 85, respectively (both S47C and L85C derived from the protein in which C102 had been replaced by threonine). All three spin-labeled protein derivatives were characterized using electron paramagnetic resonance (EPR) techniques. The continuous wave (CW) EPR spectrum of spin label attached to L85C differed from those recorded for spin label attached to C102 or S47C, indicating that spin label at position 85 was more immobilized and exhibited more complex tumbling than spin label at two other positions. The temperature dependence of the CW EPR spectra and CW EPR power saturation revealed further differences of spin-labeled L85C. The results were discussed in terms of application of the site-directed spin labeling technique in probing the local dynamic structure of iso-1-cytochrome c.
Źródło:
Acta Biochimica Polonica; 1999, 46, 4; 889-899
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
    Wyświetlanie 1-2 z 2

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