Volume 8 Supplement 1
Structure of the xenotropic murine leukaemia virus-related virus matrix protein
© Doležal et al; licensee BioMed Central Ltd. 2011
Published: 6 June 2011
We present the preparation of the xenotropic murine leukaemia virus-related virus matrix protein (XMRV-MA) and its structure determined by NMR spectroscopy.
The DNA fragment encoding XMRV-MA was obtained from prostate tumour cell cDNA (Rv1 cell line) by PCR and inserted into a pET-22b plasmid. Non-myristoylated, uniformly 13C- and 15N-labeled XMRV-MA, fused with histidine tag, was produced in E. coli BL21 (DE3) cells. The protein was purified by immobilized metal affinity chromatography (NiNTA-agarose) and size-exclusion chromatography (Sephadex 75), and then concentrated to 5 mg/ml.
All NMR data were collected at 298 K on a 600 MHz Bruker Avance III spectrometer equipped with a cryogenic triple-resonance probe and analyzed with CcpNmr Analysis. Back-bone and side-chain resonances were assigned using standard NMR experiments and structural constraints were obtained from 13C- and 15N-edited NOESY experiments. Structures were calculated with ARIA.
Although the protein sequence of the XMRV-MA is very similar to that of the murine leukaemia virus matrix protein (MLV-MA), it varies in several amino acid residues. We compared the structures of the XMRV-MA and MLV-MA and found that those changes are localized in a few domains, mostly on the surface of the protein.
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.