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Monitoring of three distinct structures of restriction enzyme complexes using characteristic fluorescence from site-selectively incorporated solvatochromic probe.
- Source :
-
Photochemical & Photobiological Sciences . Aug2007, Vol. 6 Issue 8, p836-841. 6p. - Publication Year :
- 2007
-
Abstract
- The local change in the three different structures of restriction enzyme BamHI, which include DNA-free dimer and non-specific and specific complexes with DNA, were detected by the fluorescence from a site-selectively introduced solvatochromic fluorophore Nβ-l-alanyl-5-(N,N-dimethylamino)naphthalene-1-sulfonamide (DanAla). According to the crystal structure, α-helices of the non-specific complex containing Ile82, Glu86 and Trp206 residues are converted into random coil by the formation of specific complex with a substrate. To understand the microenvironmental change caused by the structural transition around these positions, the DanAla probe was site-specifically introduced into the positions, and steady-state and time-resolved fluorescence was observed. The steady-state fluorescence gave us information that the rigidity of the polypeptide chains would be enhanced by the formation of the specific complex. The time-resolved fluorescence supported that the change in a water molecule-accessible space was induced by DNA-binding. We revealed that the change in rigidity and solvation around the specific positions was detected by the characteristic fluorescence using the combination of steady-state and time-resolved fluorescence techniques. [ABSTRACT FROM AUTHOR]
- Subjects :
- *ENZYMES
*FLUORESCENCE
*DNA
*NAPHTHALENE
Subjects
Details
- Language :
- English
- ISSN :
- 1474905X
- Volume :
- 6
- Issue :
- 8
- Database :
- Academic Search Index
- Journal :
- Photochemical & Photobiological Sciences
- Publication Type :
- Academic Journal
- Accession number :
- 26095500
- Full Text :
- https://doi.org/10.1039/b705049f