PMID- 28465237
OWN - NLM
STAT- MEDLINE
DCOM- 20171002
LR  - 20171128
IS  - 1090-2104 (Electronic)
IS  - 0006-291X (Linking)
VI  - 487
IP  - 4
DP  - 2017 Jun 10
TI  - Biochemical and functional characterization of MRA_1571 of Mycobacterium 
      tuberculosis H37Ra and effect of its down-regulation on survival in macrophages.
PG  - 892-897
LID - S0006-291X(17)30838-0 [pii]
LID - 10.1016/j.bbrc.2017.04.149 [doi]
AB  - Amino acid biosynthesis has emerged as a source of new drug targets as many 
      bacterial strains auxotrophic for amino acids fail to proliferate under in vivo 
      conditions. Branch chain amino acids (BCAAs) are important for Mycobacterium 
      tuberculosis (Mtb) survival and strains deficient in their biosynthesis were 
      attenuated for growth in mice. Threonine dehydratase (IlvA) is a 
      pyridoxal-5-phosphate (PLP) dependent enzyme that catalyzes the first step in 
      isoleucine biosynthesis. The MRA_1571 of Mycobacterium tuberculosis H37Ra 
      (Mtb-Ra), annotated to be coding for IlvA, was cloned, expressed and purified. 
      Purified protein was subsequently used for developing enzyme assay and to study 
      its biochemical properties. Also, E. coli BL21 (DE3) IlvA knockout 
      (E. coli-DeltailvA) was developed and genetically complemented with Mtb-Ra ilvA 
      expression construct (pET32a-ilvA) to make complemented E. coli strain 
      (E. coli-DeltailvA + pET32a-ilvA). The E. coli-DeltailvA showed growth failure in minimal 
      medium but growth restoration was observed in E. coli-DeltailvA + pET32a-ilvA. 
      E. coli-DeltailvA growth was also restored in the presence of isoleucine. The IlvA 
      localization studies detected its distribution in cell wall and membrane 
      fractions with relatively minor presence in cytosolic fraction. Maximum IlvA 
      expression was observed at 72 h in wild-type (WT) Mtb-Ra infecting macrophages. 
      Also, Mtb-Ra IlvA knockdown (KD) showed reduced survival in macrophages compared 
      to WT and complemented strain (KDC).
CI  - Copyright (c) 2017 Elsevier Inc. All rights reserved.
FAU - Sharma, Rishabh
AU  - Sharma R
AD  - Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 
      10, Jankipuram Extension, Sitapur Road, Lucknow - 226031, India.
FAU - Keshari, Deepa
AU  - Keshari D
AD  - Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 
      10, Jankipuram Extension, Sitapur Road, Lucknow - 226031, India.
FAU - Singh, Kumar Sachin
AU  - Singh KS
AD  - Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 
      10, Jankipuram Extension, Sitapur Road, Lucknow - 226031, India.
FAU - Singh, Sudheer Kumar
AU  - Singh SK
AD  - Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 
      10, Jankipuram Extension, Sitapur Road, Lucknow - 226031, India; Academy of 
      Scientific and Industrial Research (AcSIR), New Delhi, India. Electronic address: 
      sudheer_singh@cdri.res.in.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170502
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (Bacterial Proteins)
RN  - EC 4.3.1.19 (Threonine Dehydratase)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/chemistry/*metabolism
MH  - Cells, Cultured
MH  - *Down-Regulation
MH  - Macrophages/*metabolism/*microbiology
MH  - Mice
MH  - Mycobacterium tuberculosis/*enzymology/*metabolism
MH  - Threonine Dehydratase/chemistry/*metabolism
OTO - NOTNLM
OT  - Biochemical and functional characterization
OT  - Localization
OT  - Macrophages
OT  - Mycobacterium tuberculosis
OT  - Threonine dehydratase
EDAT- 2017/05/04 06:00
MHDA- 2017/10/03 06:00
CRDT- 2017/05/04 06:00
PHST- 2017/04/25 00:00 [received]
PHST- 2017/04/29 00:00 [accepted]
PHST- 2017/05/04 06:00 [pubmed]
PHST- 2017/10/03 06:00 [medline]
PHST- 2017/05/04 06:00 [entrez]
AID - S0006-291X(17)30838-0 [pii]
AID - 10.1016/j.bbrc.2017.04.149 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 2017 Jun 10;487(4):892-897. doi: 
      10.1016/j.bbrc.2017.04.149. Epub 2017 May 2.