6kl0
Crystal structure of the S65T/F99S/M153T/V163A variant of perdeuterated GFP at pD 7.0Crystal structure of the S65T/F99S/M153T/V163A variant of perdeuterated GFP at pD 7.0
Structural highlights
FunctionGFP_AEQVI Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Publication Abstract from PubMedHydrogen atoms are critical to the nature and properties of proteins, and thus deuteration has the potential to influence protein function. In fact, it has been reported that some deuterated proteins show different physical and chemical properties to their protiated counterparts. Consequently, it is important to investigate protonation states around the active site when using deuterated proteins. Here, hydrogen isotope effects on the S65T/F99S/M153T/V163A variant of green fluorescent protein (GFP), in which the deprotonated B form is dominant at pH 8.5, were investigated. The pH/pD dependence of the absorption and fluorescence spectra indicates that the protonation state of the chromophore is the same in protiated GFP in H(2)O and protiated GFP in D(2)O at pH/pD 8.5, while the pK(a) of the chromophore became higher in D(2)O. Indeed, X-ray crystallographic analyses at sub-angstrom resolution revealed no apparent changes in the protonation state of the chromophore between the two samples. However, detailed comparisons of the hydrogen OMIT maps revealed that the protonation state of His148 in the vicinity of the chromophore differed between the two samples. This indicates that protonation states around the active site should be carefully adjusted to be the same as those of the protiated protein when neutron crystallographic analyses of proteins are performed. X-ray crystallographic studies on the hydrogen isotope effects of green fluorescent protein at sub-angstrom resolutions.,Tai Y, Takaba K, Hanazono Y, Dao HA, Miki K, Takeda K Acta Crystallogr D Struct Biol. 2019 Dec 1;75(Pt 12):1096-1106. doi: , 10.1107/S2059798319014608. Epub 2019 Nov 19. PMID:31793903[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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