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PhD | Biomoléculas, metaloproteínas, electron transfer ⚡👩‍🔬 | Feminista 💚 | Futbolista ⚽️ | Arg 🇦🇷 | She/Her |

Sep 7, 2020, 7 tweets

Hello @LatinXChem! Today I want to share my latest work about the mechanisms of assembly of CuA Site by mitochondrial metallochaperones. Done at @IBR_CONICET and @VilaLab, in collaboration with @soy_MAC and @labriataphd. #LatinXChem #LatinXChemBio #BIO045 (1/5)

We found that A. thaliana Sco1 binds both Cu(I) and Cu(II) with high affinity. We analyzed both copper sites by spectroscopic methods, XRay crystallograpy and DFT, confirming that Cu(I) tri-coordinated (2SCys-NeHis) and Cu(II) tetra-coordinated (2SCys-NeHis-H2O) (2/5)

To test the activity of AtSco1, we first designed a soluble model of AtCoxII. We confirmed the formation of CuA site and the possibility of monitoring all four possible oxidation and metallation states of the protein by NMR. (3/5)

We tested the Cu(II) metallochaperone activity of AtSco1 using the model of AtCoxII. Cu(II)Sco:CoxII* at 1:1 ratio forms a pink adduct (stable + 48 h). If we add a 2nd eq of ScoCu(II), the color turns unstable, fully disappearing after 12h. Reduced CuA is formed. (4/5)

We analyzed the perturbed residues in the NMR spectra when the pink adduct is formed. Based on this information, we modeled the interaction between the two proteins. (5/5)

We acknowledge @IBR_CONICET for being an amazing place to do science ♥ and @CONICETDialoga for support !

I can't believe I @ the wrong Lab! haha the work was done at @lab_vila 😱😅

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