ISSN Print 2500–1094
ISSN Online 2542–1204
Bulletin of RSMU
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
ORIGINAL RESEARCH
Two HMG domains of yeast mitochondrial protein Abf2p have different affinity to DNA
About authors
1 Department of Molecular Biology, Faculty of Biology,Lomonosov Moscow State University, Moscow, Russia
2 Department of General Surgery, Faculty of Fundamental Medicine,Lomonosov Moscow State University, Moscow, Russia
Correspondence should be addressed: Peter Kamenski
Leninskie gory, d. 1, str. 12, Moscow, Russia, 119991; moc.liamg@iksnemak.rtoip
About paper
Funding: this study was supported by the Russian Foundation for Basic Research (project no. 14-04-31554 mol_a).
Received: 2015-09-29
Accepted: 2015-12-09
Published online: 2017-01-05
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Fig. 1. Isolation and purification of recombinant proteins corresponding to HMG1 and HMG2 domains of the Abf2p protein
Recombinant proteins were purified by metal affinity chromatography on Ni sepharose. 1 — molecular weight markers (the molecular weights of marker proteins are shown on the left); 2, 6 — damaged cell lysates; 3, 7 — factions that didn’t interact with affinity column; 4, 8 — purified preparations of recombinant proteins HMG1 and HMG2 respectively.
Fig. 2. Analysis of the binding of recombinant proteins corresponding to HMG1 and HMG2 domains with DNA using EMSA
Linear DNA duplex (double-stranded DNA - dsDNA) and cruciform DNA (4-way junction — 4wj) at a concentration of 10 nM were incubated with increasing concentrations of recombinant proteins, after which the reaction mixtures were separated in 6 % polyacrylamide gel. (A) Binding with DNA of HMG1 domain; 1–4 — interaction with DNA duplex, 5–9 — binding with cruciform DNA; C1 — resulting complex. (B) Binding with DNA of HMG2 domain; 1–3 — interaction with DNA duplex, 4–6 — binding with cruciform DNA. The lower part of the figure shows the used concentration of recombinant proteins (in nM).
