Resonance assignments and secondary structure prediction of secretory protein Rv0603 from Mycobacterium tuberculosis H37
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Resonance assignments and secondary structure prediction of secretory protein Rv0603 from Mycobacterium tuberculosis H37Rv Sarita Tripathi1,2 · Rahul Yadav1 · Anupam Jain1 · Surya V. S. R. K. Pulavarti1 · Prem Prakash Pathak1 · Ajaya Kumar Behera2 · Ashish Arora1 Received: 2 March 2020 / Accepted: 14 May 2020 © Springer Nature B.V. 2020
Abstract We report the NMR resonance assignments of N-terminal signal sequence deleted secretory protein Rv0603 (∆1–28−Rv0603) from Mycobacterium tuberculosis H37Rv. ∆1–28−Rv0603 displayed good peak yield and signal dispersion in 2D [ 15N-1H] HSQC spectrum, which prompted us to proceed for resonance assignments on this construct. Standard triple-resonance experiments for resonance assignments were recorded on [U-15N]-∆Rv0603 and [U-15N, 13C]-∆Rv0603 samples. We obtained 97% of backbone 1HN, 98% of 13Cα, 98% of 1Hα, 96% of 13C´, 100% of 13Cβ, 100% of 1Hβ and 98% of side-chain 1H chemical shifts. This protein does not show any sequence similarity to any other protein of known structure. Determination of its solution structure would facilitate understanding of its biological function. Keywords ∆1–28−Rv0603 · Secreted protein · NMR · Resonance assignment
Biological context Mycobacterium tuberculosis secretes a variety of proteins into its surrounding environment, many of which play an important role in host–pathogen interaction. Rv0603 was identified as a Sec-dependent secreted protein that is specific to the M. tuberculosis Complex group of pathogens, apart from which it is present only in M. kansasii (Amor et al. 2005). Intriguingly, Rv0603 displayed reactivity towards the serum derived from untreated TB patients as well as the patients having respiratory diseases other than tuberculosis (Amor et al. 2005). M. tuberculosis H37Rv Rv0603 is encoded as a 103-residue protein, including the N-terminal stretch of residues 1–28, which have been identified by the SignalP program (Gomez et al. 2000) to be a signal peptide for Sec-dependent protein secretion pathway (Gomez et al. 2000, Chiliza et. al. 2017).
* Ashish Arora [email protected] 1
Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226 031, India
School of Chemistry, Sambalpur University, Jyoti Vihar, Sambalpur, Odisha 768019, India
2
So far, there is no significant sequence similarity of Rv0603 with any protein with known three-dimensional structure or function. Although, initially annotated as a hypothetical protein, Rv0603 has later been characterized as a protein associated with cell wall and cell processes (Becq et. al. 2007). The ORF rv0603 excluding nucleotides encoding signal peptide was cloned and expressed, yielding the exported form of Rv0603, Δ1-28−Rv0603. As part of our efforts in understanding the role of secretory proteins in the pathogenicity of M. tuberculosis, we have undertaken structural characterization of M. tuberculosis Δ1-28−Rv0603 by solution NMR spectroscopy. Here, we report the expression, purification, backbone and side-cha
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