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Preparation of the recombinant НU-proteins from S. melliferum and M. gallisepticum and of their complexes with DS-DNA for structural NMR experiments

We created a high-performance system for expression of 13 C, 15 N-double-labeled recombinant НU-proteins from mycoplasms S. melliferum (HUSpm) and M. gallisepticum (HUMgal), and a purification protocol that yields protein samples suitable for structure studies by high-resolution NMR. Using these pro...

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Bibliographic Details
Published in:Moscow University chemistry bulletin 2016-07, Vol.71 (4), p.221-226
Main Authors: Altukhov, D. A., Agapova, Y. K., Vlaskina, A. V., Korzhenevskiy, D. A., Nikolaeva, A. Y., Frank-Kamenetskaya, A. M., Bocharov, E. V., Rakitina, T. V.
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Language:English
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Summary:We created a high-performance system for expression of 13 C, 15 N-double-labeled recombinant НU-proteins from mycoplasms S. melliferum (HUSpm) and M. gallisepticum (HUMgal), and a purification protocol that yields protein samples suitable for structure studies by high-resolution NMR. Using these proteins we optimized the conditions of NMR experiments and obtained two-dimensional heteronuclear 1 H/ 15 N-HSQC and 1 H/ 13 С-HSQC-CT NMR spectra of free НU-proteins (HUSpm and HUMGal), and 1H/15N-HSQC spectrum of a complex between HUSpm and a double-stranded DNA (ds-DNA). The techniques we created are applicable in the studies of НU-proteins from other organisms. They allow analyzing structure and dynamics of free proteins and of their conformational changes induced by binding of various ds-DNAs or of DNA binding inhibitors.
ISSN:0027-1314
1935-0260
DOI:10.3103/S0027131416040027