IRFI 東京工業大学国際先駆研究機構 IRFI 東京工業大学国際先駆研究機構



[Paper] “Lanmodulin peptides – unravelling the binding of the EF-Hand loop sequences stripped from the structural corset” published by WRH professor Satoru Tsushima

WRH professor Satoru Tsushima (IIR Laboratory for Zero-Carbon Energy) published a paper in Inorganic Chemistry Frontiers (with illustration on inside front cover).

“Lanmodulin peptides – unravelling the binding of the EF-Hand loop sequences stripped from the structural corset”

Lanmodulin (LanM), a naturally lanthanide (Ln)-binding protein with a remarkable selectivity for Lns over Ca(II) and affinities in the picomolar range, is an attractive target to address challenges in Ln separation. Why LanM has such a high selectivity is currently not entirely understood; both specific amino acid sequences of the EF-Hand loops and cooperativity effects have been suggested. Here, we removed the effect of cooperativity and synthesised all four 12-amino acid EF-Hand loop peptides, and investigated their affinity for two Lns (Eu(III) and Tb(III)), the actinide Cm(III) and Ca(II). Using isothermal titration calorimetry and time-resolved laser fluorescence spectroscopy (TRLFS) combined with parallel factor analysis, we show that the four short peptides behave very similarly, having affinities in the micromolar range for Eu(III) and Tb(III). Ca(II) was shown not to bind to the peptides, which was verified with circular dichroism spectroscopy. This technique also revealed an increase in structural organisation upon Eu(III) addition, which was supported by molecular dynamics simulations. Lastly, we put Eu(III) and Cm(III) in direct competition using TRLFS. Remarkably, a slightly higher affinity for Cm(III) was found. Our results demonstrate that the picomolar affinities in LanM are largely an effect of pre-structuring and therefore a reduction of flexibility in combination with cooperative effects, and that all EF-Hand loops possess similar affinities when detached from the protein backbone, albeit still retaining the high selectivity for lanthanides and actinides over calcium.



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