Fluorescence quenching of buried Trp residues by acrylamide does not require penetration of the protein fold

The Journal of Physical Chemistry. B
Giovanni B Strambini, Margherita Gonnelli

Abstract

The accessibility of acrylamide to buried Trp residues in proteins, as attested by dynamic quenching of their fluorescence emission, is often interpreted in terms of migration of the quencher (Q) through the globular fold. The quencher penetration mechanism, however, has long been debated because, on one hand, solutes the size of acrylamide are not expected to diffuse within the protein matrix on the nanosecond time scale of fluorescence and, on the other hand, alternative reactions pathways where Q remains in the solvent cannot be ruled out. To test the Q penetration hypothesis, we compared the quenching rates of acrylamide analogs of increasing molecular size (acrylonitrile, acrylamide, and bis-acrylamide) on the buried Trp residues of RNaseT1 and parvalbumin. The results show that the largest molecule, bis-acrylamide, is also the most efficient quencher and that in general the quenching rate is not correlated to quencher size, as expected for a penetration mechanism. Whereas these results rule out significant internal Q migration in the times of fluorescence, it is also demonstrated that up to a depth of burial of 3 A, through-space interactions with acrylamide in the solvent satisfactorily account for the small rate constan...Continue Reading

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Citations

Sep 19, 2012·Journal of Pharmaceutical Sciences·Keith A BrittTheodore W Randolph
Dec 25, 2015·The Journal of Physical Chemistry Letters·Nirbhik AcharyaSantosh Kumar Jha
Nov 26, 2013·Biophysical Journal·Saswata Sankar SarkarGuruswamy Krishnamoorthy
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Aug 2, 2017·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Caitlin J Jenvey, Judith R Stabel
Feb 7, 2019·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Sasi DivyaDeepa Khushalani
May 10, 2019·PloS One·Leina Moro PérezRancés Blanco Santana
Jul 16, 2019·International Journal of Biological Macromolecules·P Prasanthan, N Kishore
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Aug 14, 2019·The Journal of Physical Chemistry. B·Anu KadyanSiddharth Pandey

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