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

Autor:	Alena Yu. Nikolaeva, D. A. Altukhov, E. V. Bocharov, D.A. Korzhenevskiy, Yuliya K. Agapova, Tatiana V. Rakitina, Anna V. Vlaskina, A. M. Frank-Kamenetskaya
Rok vydání:	2016
Předmět:	0301 basic medicine Chemistry Stereochemistry 030106 microbiology General Chemistry law.invention NMR spectra database 03 medical and health sciences Crystallography chemistry.chemical_compound 030104 developmental biology Heteronuclear molecule law Recombinant DNA Gene DNA
Zdroj:	Moscow University Chemistry Bulletin. 71:221-226
ISSN:	1935-0260 0027-1314
Popis:	We created a high-performance system for expression of 13C, 15N-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 1H/15N-HSQC and 1H/13С-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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::61df861f229cbfe063b5ef69a457ab62 https://doi.org/10.3103/s0027131416040027 Zobrazit plný text záznamu Full text from SpringerLink