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1.
J Fluoresc ; 26(1): 113-9, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26482593

RESUMEN

Two acyl hydrazone derivatives, AHI and AHN,made from ibuprofen and naproxen-derived hydrazides, were prepared and studied of binding properties with serine protease trypsin by UV-vis absorption and fluorescence quenching at pH 7.4. The results suggest that both hydrazones can interact strongly with trypsin and there are the formation of trypsin-hydrazone complexes. The Stern-Volmer constants, binding constants,binding sites and the corresponding thermodynamic parameters ΔH°, ΔS° and ΔG° were calculated at different temperatures.The effect of common metal ions on the constants was also discussed. The binding modes can be explained on the basis of hydrogen bonds and van der Waals forces. The binding distance(r) ~3 nm between the donor (trypsin) and acceptors (AHI and AHN) was obtained according to Förster's non-radiative energy transfer theory. Moreover, LOD and LOQ of hydrazones were calculated in the presence of trypsin.


Asunto(s)
Hidrazonas/química , Ibuprofeno/química , Naproxeno/química , Tripsina/química , Sitios de Unión , Hidrazonas/síntesis química , Estructura Molecular , Espectrometría de Fluorescencia , Termodinámica , Tripsina/metabolismo
2.
PLoS One ; 9(9): e105751, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25244619

RESUMEN

The Escherichia coli protein BdcA (previously referred to as YjgI) plays a key role in the dispersal of cells from bacterial biofilms, and its constitutive activation provides an attractive therapeutic target for dismantling these communities. In order to investigate the function of BdcA at a molecular level, we integrated structural and functional studies. Our 2.05 Å structure of BdcA shows that it is a member of the NAD(P)(H)-dependent short-chain dehydrogenase/reductase (SDR) superfamily. Structural comparisons with other members of the SDR family suggested that BdcA binds NADP(H). This was demonstrated experimentally using thermal denaturation studies, which showed that BcdA binds specifically to NADPH. Subsequent ITC experiments further confirmed this result and reported a Kd of 25.9 µM. Thus, BdcA represents the newest member of the limited number of oxidoreductases shown to be involved in quorum sensing and biofilm dispersal.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , NADP/metabolismo , Unión Proteica , Conformación Proteica , Especificidad por Sustrato
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