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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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Published in: | Moscow University chemistry bulletin 2016-07, Vol.71 (4), p.221-226 |
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Main Authors: | , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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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. |
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ISSN: | 0027-1314 1935-0260 |
DOI: | 10.3103/S0027131416040027 |