PMID- 30615070
OWN - NLM
STAT- MEDLINE
DCOM- 20190624
LR  - 20190624
IS  - 1745-7270 (Electronic)
IS  - 1672-9145 (Linking)
VI  - 51
IP  - 2
DP  - 2019 Feb 1
TI  - Biophysical characterization and ligand-binding properties of the elongation 
      factor Tu from Mycobacterium tuberculosis.
PG  - 139-149
LID - 10.1093/abbs/gmy164 [doi]
AB  - Mycobacterium tuberculosis (Mtb) is the key devastating bacterial pathogen 
      responsible for tuberculosis. Increasing emergence of multi-drug-resistant, 
      extensively drug-resistant, and rifampicin/isoniazid-resistant strains of Mtb 
      makes the discovery of validated drug targets an urgent priority. As a vital 
      translational component of the protein biosynthesis system, elongation factor Tu 
      (EF-Tu) is an important molecular switch responsible for selection and binding of 
      the cognate aminoacyl-tRNA to the acceptor site on the ribosome. In addition, 
      EF-Tu from Mtb (MtbEF-Tu) is involved in the initial step of trans-translation 
      which is an effective system for rescuing the stalled ribosomes from non-stop 
      translation complexes under stress conditions. Given its crucial role in protein 
      biosynthesis, EF-Tu is identified as an excellent molecular target for drug 
      design. Here, we reported the recombinant expression, purification, biophysical 
      characterization, and structural modeling of the MtbEF-Tu protein. Our results 
      demonstrated that prokaryotic expression plasmids of pET28a-MtbEF-Tu could be 
      expressed efficiently in Escherichia coli. We successfully purified the 6x 
      His-tagged proteins with a yield of 16.8 mg from 1 l of Luria Bertani medium. 
      Dynamic light scattering experiments showed that MtbEF-Tu existed in a monomeric 
      form, and circular dichroism experiments indicated that MtbEF-Tu was well 
      structured. Moreover, isothermal titration calorimetry experiments displayed that 
      the purified MtbEF-Tu protein possessed intermediate binding affinities for 
      guanosine-5'-triphosphate (GTP) and GDP. The GTP/GDP-binding sites were predicted 
      by flexible molecular docking approach which reveals that GTP/GDP binds to 
      MtbEF-Tu mainly through hydrogen bonds. Our work lays the essential basis for 
      further structural and functional studies of MtbEF-Tu as well as MtbEF-Tu-related 
      novel drug developments.
CI  - (c) The Author(s) 2019. Published by Oxford University Press on behalf of the 
      Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological 
      Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, 
      please e-mail: journals.permissions@oup.com.
FAU - Yang, Juanjuan
AU  - Yang J
AD  - Institute of Pharmaceutical Biotechnology and Engineering, College of Biological 
      Science and Biotechnology, Fuzhou University, Fuzhou, China.
FAU - Hong, Jing
AU  - Hong J
AD  - Institute of Pharmaceutical Biotechnology and Engineering, College of Biological 
      Science and Biotechnology, Fuzhou University, Fuzhou, China.
FAU - Luo, Ling
AU  - Luo L
AD  - Institute of Pharmaceutical Biotechnology and Engineering, College of Biological 
      Science and Biotechnology, Fuzhou University, Fuzhou, China.
FAU - Liu, Ke
AU  - Liu K
AD  - State Key Laboratory of Stress Cell Biology, School of Life Sciences, Xiamen 
      University, Xiamen, China.
FAU - Meng, Chun
AU  - Meng C
AD  - Institute of Pharmaceutical Biotechnology and Engineering, College of Biological 
      Science and Biotechnology, Fuzhou University, Fuzhou, China.
FAU - Ji, Zhi-Liang
AU  - Ji ZL
AD  - State Key Laboratory of Stress Cell Biology, School of Life Sciences, Xiamen 
      University, Xiamen, China.
FAU - Lin, Donghai
AU  - Lin D
AD  - High-Field NMR Center, Key Laboratory for Chemical Biology of Fujian Province, 
      College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, China.
LA  - eng
PT  - Journal Article
PL  - China
TA  - Acta Biochim Biophys Sin (Shanghai)
JT  - Acta biochimica et biophysica Sinica
JID - 101206716
RN  - 0 (Bacterial Proteins)
RN  - 0 (Ligands)
RN  - 0 (RNA, Transfer, Amino Acyl)
RN  - 146-91-8 (Guanosine Diphosphate)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - EC 3.6.1.- (Peptide Elongation Factor Tu)
SB  - IM
MH  - Bacterial Proteins/chemistry/genetics/*metabolism
MH  - Binding Sites
MH  - Guanosine Diphosphate/chemistry/metabolism
MH  - Guanosine Triphosphate/chemistry/metabolism
MH  - Ligands
MH  - Models, Molecular
MH  - Mutation
MH  - Mycobacterium tuberculosis/genetics/*metabolism
MH  - Peptide Elongation Factor Tu/chemistry/genetics/*metabolism
MH  - Protein Binding
MH  - *Protein Biosynthesis
MH  - Protein Domains
MH  - RNA, Transfer, Amino Acyl/metabolism
MH  - Ribosomes/metabolism
OTO - NOTNLM
OT  - MtbEF-Tu
OT  - protein biosynthesis
OT  - protein expression and purification
OT  - protein-guanine nucleotide interaction
OT  - tuberculosis
EDAT- 2019/01/08 06:00
MHDA- 2019/06/25 06:00
CRDT- 2019/01/08 06:00
PHST- 2018/07/27 00:00 [received]
PHST- 2018/11/28 00:00 [revised]
PHST- 2018/11/30 00:00 [accepted]
PHST- 2019/01/08 06:00 [pubmed]
PHST- 2019/06/25 06:00 [medline]
PHST- 2019/01/08 06:00 [entrez]
AID - 5274658 [pii]
AID - 10.1093/abbs/gmy164 [doi]
PST - ppublish
SO  - Acta Biochim Biophys Sin (Shanghai). 2019 Feb 1;51(2):139-149. doi: 
      10.1093/abbs/gmy164.