PMID- 24266429
OWN - NLM
STAT- MEDLINE
DCOM- 20140409
LR  - 20140214
IS  - 1362-3095 (Electronic)
IS  - 0955-3002 (Linking)
VI  - 90
IP  - 2
DP  - 2014 Feb
TI  - Novel method for screening of radioprotective agents providing protection to DNA 
      ligase against gamma radiation induced damage.
PG  - 187-92
LID - 10.3109/09553002.2014.868613 [doi]
AB  - PURPOSE: A simple, sensitive and novel method was developed to screen out 
      potential agents able to protect functional activity of DNA ligase against gamma 
      irradiation-induced damage. Repeatability, authenticity and sensitivity of the 
      method was verified by analyzing DNA ligase protecting activities of well-known 
      radioprotectors such as amifostine, trolox, melatonin, semiquinone glucoside 
      derivative (SQGD) and an antioxidant gallic acid in extremely low concentration 
      (1 mug/reaction). MATERIAL AND METHODS: Two different sets (Set A and B) of T4 DNA 
      ligase (1 unit/set) were prepared. Set 'A' (negative control) was exposed to 
      different doses (3-5 kGy) of gamma radiation in the absence of radioprotective 
      compounds. Set B (test) was exposed to similar doses of gamma radiation in the 
      presence of radioprotective compounds. Following irradiation, DNA ligase was 
      mixed with lambda DNA (250 ng) pre-digested with Hind III restriction endonuclease. 
      Ligation reaction was performed in both sets simultaneously at 22 degrees C for 20 min 
      and reaction product was analyzed using agarose gel electrophoresis. RESULTS: 
      Complete DNA ligation was observed in samples where DNA ligase was irradiated in 
      the presence of radioprotectective compounds, i.e., amifostine, trolox, melatonin 
      and a natural radioprotector semiquinone glucoside derivative (SQGD) 
      individually, while, functional impairment in ligation activity of DNA ligase was 
      evident in samples in which DNA ligase was irradiated in the absence of a 
      radioprotective compound. CONCLUSION: The current method was able to provide 
      significant input to screen out radioprotective compounds able to protect DNA 
      ligase functional activity against gamma radiation-induced functional impairment.
FAU - Mishra, Saurabh
AU  - Mishra S
AD  - Radiation Biotechnology Laboratory, Department of Radiation Biosciences.
FAU - Malhotra, Poonam
AU  - Malhotra P
FAU - Gupta, Ashutosh K
AU  - Gupta AK
FAU - Singh, Praveen K
AU  - Singh PK
FAU - Mishra, Anil K
AU  - Mishra AK
FAU - Javed, Saleem
AU  - Javed S
FAU - Kumar, Raj
AU  - Kumar R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140109
PL  - England
TA  - Int J Radiat Biol
JT  - International journal of radiation biology
JID - 8809243
RN  - 0 (Radiation-Protective Agents)
RN  - EC 6.5.1.- (DNA Ligases)
SB  - IM
MH  - DNA Ligases/physiology/*radiation effects
MH  - Drug Evaluation, Preclinical/*methods
MH  - *Gamma Rays
MH  - Radiation-Protective Agents/*pharmacology
EDAT- 2013/11/26 06:00
MHDA- 2014/04/10 06:00
CRDT- 2013/11/26 06:00
PHST- 2013/11/26 06:00 [entrez]
PHST- 2013/11/26 06:00 [pubmed]
PHST- 2014/04/10 06:00 [medline]
AID - 10.3109/09553002.2014.868613 [doi]
PST - ppublish
SO  - Int J Radiat Biol. 2014 Feb;90(2):187-92. doi: 10.3109/09553002.2014.868613. Epub 
      2014 Jan 9.