biotin label
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2018 ◽  
Vol 23 (6) ◽  
pp. 545-553 ◽  
Author(s):  
Ming Ji ◽  
Liyuan Wang ◽  
Nina Xue ◽  
Fangfang Lai ◽  
Sen Zhang ◽  
...  

Poly(ADP-ribose) polymerase 3 (PARP3) is an important member of the PARP family and shares high structural similarities with both PARP1 and PARP2. The biological roles of PARP3 are currently under investigation; however, several key reports indicate the integral roles of PARP3 in DNA damage repair, and thus it has been investigated as a novel target in oncology. It is clear that the identification of selective PARP3 inhibitors would further advance the understanding of the biological roles of PARP3. Herein, we describe a modified PARP3 screening assay using biotinylated NAD+ as the specialized substrate. This method relies on the activity of PARP3 to transfer the biotinylated NAD+ onto a histone protein to form ADP-ribosylated histone. The biotin label on this histone protein is then detected and quantifies the intrinsic enzymatic activity of PARP3. We optimized the assay with respect to the histone, NAD+/biotinylated NAD+ mixture, DNA, and PARP3. Our developed screening system was then validated with a reported selective PARP3 inhibitor, ME0328, as well as utilizing five other clinically available PARP1/2 inhibitors. We demonstrated that our assay system was sensitive, efficient, and economical, and we reason that it could be useful for the development of highly selective PARP3 inhibitors in the future.


2014 ◽  
Vol 28 (3) ◽  
pp. 114-125 ◽  
Author(s):  
Donald M. Mock ◽  
John A. Widness ◽  
Peter Veng-Pedersen ◽  
Ronald G. Strauss ◽  
Jose A. Cancelas ◽  
...  

2013 ◽  
Vol 31 (3) ◽  
pp. 1277-1281 ◽  
Author(s):  
HISAAKI MIYOSHI ◽  
ASAHIRO MORISHITA ◽  
JOJI TANI ◽  
TEPPEI SAKAMOTO ◽  
KOJI FUJITA ◽  
...  

Transfusion ◽  
2011 ◽  
Vol 52 (5) ◽  
pp. 1097-1105 ◽  
Author(s):  
Donald M. Mock ◽  
Gary L. Lankford ◽  
Nell I. Matthews ◽  
Leon F. Burmeister ◽  
Daniel Kahn ◽  
...  

Author(s):  
Ruochun Huang ◽  
Weidong Jiang ◽  
Jean Yang ◽  
Ying Qing Mao ◽  
Ying Zhang ◽  
...  

2009 ◽  
Vol 23 (S1) ◽  
Author(s):  
Nell I Matthews ◽  
Donald M Mock ◽  
Ronald G Strauss ◽  
Leon F Burmeister ◽  
Robert Schmidt ◽  
...  

2009 ◽  
Vol 23 (S1) ◽  
Author(s):  
Nell I Matthews ◽  
Donald M Mock ◽  
Ronald G Strauss ◽  
Leon F Burmeister ◽  
Robert Schmidt ◽  
...  

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