Illuminating allosteric conformational change with an environmentally sensitive fluorescent probe

2015 ◽  
Vol 2015 (1) ◽  
pp. c1-4
Author(s):  
Robert B. Freedman ◽  
Alan D.B. Malcolm
Keyword(s):  
Fluorescent Probe ◽  
Conformational Change ◽  
Environmentally Sensitive

scholarly journals Allosteric regulation: Illuminating allosteric conformational change with an environmentally sensitive fluorescent probe

2015 ◽  
Vol 37 (3) ◽  
pp. 54-57
Author(s):  
Robert B. Freedman ◽  
Alan D.B. Malcolm
Keyword(s):  
Glutamate Dehydrogenase ◽  
Fluorescent Probe ◽  
Conformational Change ◽  
Conformational Changes ◽  
Allosteric Regulation ◽  
Structural Data ◽  
Biochemical Journal ◽  
Environmentally Sensitive ◽  
The 1960S ◽  
Classic Paper

Allosteric regulation was a hot topic in the 1960s, but there was very limited structural data on allosteric equilibria, and no solid information on the rates of allosteric conformational changes. In this Biochemical Journal Classic paper from 1969 George Radda and his first D.Phil. student, George Dodd determined the rate of allosteric transition in the regulatory enzyme glutamate dehydrogenase by a method new in the 1960s, the fluorescence of an environmentally sensitive extrinsic probe.

scholarly journals Conformational changes at the active site of creatine kinase at low concentrations of guanidinium chloride

1993 ◽  
Vol 291 (1) ◽  
pp. 103-107 ◽  
Author(s):  
H M Zhou ◽  
X H Zhang ◽  
Y Yin ◽  
C L Tsou
Keyword(s):  
Creatine Kinase ◽  
Fluorescent Probe ◽  
Conformational Change ◽  
Conformational Changes ◽  
Active Site ◽  
Emission Intensity ◽  
Enzyme Inactivation ◽  
Amino Groups ◽  
Guanidinium Chloride ◽  
Low Concentrations

It has been previously reported that, during denaturation of creatine kinase by guanidinium chloride (GdmCl) or urea [Tsou (1986), Trends Biochem. Sci. 11, 427-429], inactivation occurs before noticeable conformational change can be detected, and it is suggested that the conformation at the active site is more easily perturbed and hence more flexible than the molecule as a whole. In this study, the thiol and amino groups at or near the active site of creatine kinase are labelled with o-phthalaldehyde to form a fluorescent probe. Both the emission intensity and anisotropy decrease during denaturation indicating exposure of this probe and increased mobility of the active site. The above conformational changes take place together with enzyme inactivation at lower GdmCl concentrations than required to bring about intrinsic fluorescence changes of the enzyme. At the same GdmCl concentration, the rate of exposure of the probe is comparable with that of inactivation and is several orders of magnitude faster than that for the unfolding of the molecule as a whole.

The Use of 1-Anilino-8-naphthalene Sulfonate as Fluorescent Probe for Conformational Studies on Ribulose-1,5-bisphosphate Carboxylase

1976 ◽  
Vol 31 (5-6) ◽  
pp. 267-271 ◽  
Author(s):  
G. Wildner
Keyword(s):  
Carbon Dioxide ◽  
Temperature Dependence ◽  
Fluorescent Probe ◽  
Conformational Change ◽  
Temperature Effects ◽  
Fluorescence Emission ◽  
Enzyme Molecule ◽  
Bisphosphate Carboxylase ◽  
Conformational Studies

The influence of Mg2+ ions and temperature on the structure of the enzyme ribulose-1,5-bisphosphate carboxylase was investigated using the fluorescent probe 1-anilino-8-naphthalene sulfonate (ANS). The binding of ANS to the enzyme molecule caused a significant increase of fluorescence emission which was further enhanced by the addition of Mg2+. The temperature dependence of the fluorescence emission indicated a conformational change of the enzyme between 12 and 24 0C. The Mg2+ and the temperature effects were additive. ANS itself did not change the conformation of the enzyme. The influence of the substrates carbon dioxide and ribulose-1,5-bisphosphate, and the effect of the pH of the medium and of a sulfhydryl reducing reagent on fluorescence emission were analysed.

scholarly journals A fluorescent probe designed for studying protein conformational change

2005 ◽  
Vol 102 (4) ◽  
pp. 965-970 ◽  
Author(s):  
B. E. Cohen ◽  
A. Pralle ◽  
X. Yao ◽  
G. Swaminath ◽  
C. S. Gandhi ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Conformational Change ◽  
Protein Conformational Change
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