Enhancing the fluorescence of tyr-59 in ubiquitin by blocking proton transfer
Literature Information
Hana Gerbelová, Tiago Q. Faria, Maria Manuel Sampaio, Rainer Rudolph, António L. Maçanita, Helena Santos
Two highly fluorescent mutants of ubiquitin (E51Q and E51A) were produced by mutation of a single amino acid, demonstrating that excited-state proton transfer from the tyrosine residue to the carboxylate group of Glu-51 in ubiquitin is responsible for the reduced fluorescence of wild-type ubiquitin (wt-UBQ) at pH 5. E51A shows a Tm = 59 °C at pH 1.5 and a Tm > 80 °C at pH 5 similar to wt-UBQ, which shows that the mutation has not altered the protein structure significantly. The high and constant fluorescence from pH 1.5 to pH 7 allows for the study of the folding/unfolding over a wide range of pHs which would otherwise be impossible with wt-UBQ.
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