Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins
Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins
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通过高分辨率聚丙烯酰胺凝胶电泳测量聚（ADP-核糖）糖水解酶活性：组蛋白和核基质蛋白的特异性抑制
DOI：
10.1023/a:1006927119100
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发表时间：
1999
影响因子：
4.3
通讯作者：
R. Alvarez‐Gonzalez
中科院分区：
文献类型：
--
作者：
G. Pacheco‐Rodriguez;R. Alvarez‐Gonzalez
我们开发了一种新的酶分析方法，可以通过高分辨率聚丙烯酰胺凝胶电泳同时测定聚（adp -核糖）与核蛋白的非共价相互作用以及聚（adp -核糖）糖水解酶（PARG）的活性。用纯聚（adp -核糖）聚合酶（PARP）合成2 ~ 70个残基大小的adp -核糖链，并在碱性释放后用硼酸树脂亲和层析纯化。该制备的adp核糖聚合物被用作纯化PARG的酶底物。我们还获得了大鼠肝核的核基质部分，并测量了纯化的PARG在存在或不存在组蛋白或核基质蛋白的情况下的酶活性。两者都导致PARG活性的显著抑制，这是通过减少单体adp核糖的形成来确定的。PARG的抑制可能是由于这些蛋白与游离adp核糖聚合物的非共价相互作用。因此，在测量PARG活性时应考虑组蛋白和核基质蛋白的存在。
We have developed a novel enzyme assay that allows the simultaneous determination of noncovalent interactions of poly(ADP-ribose) with nuclear proteins as well as poly(ADP-ribose) glycohydrolase (PARG) activity by high resolution polyacrylamide gel electrophoresis. ADP-ribose chains between 2 and 70 residues in size were enzymatically synthesized with pure poly(ADP-ribose) polymerase (PARP) and were purified by affinity chromatography on a boronate resin following alkaline release from protein. This preparation of polymers of ADP-ribose was used as the enzyme substrate for purified PARG. We also obtained the nuclear matrix fraction from rat liver nuclei and measured the enzyme activity of purified PARG in the presence or absence of either histone proteins or nuclear matrix proteins. Both resulted in a marked inhibition of PARG activity as determined by the decrease in the formation of monomeric ADP-ribose. The inhibition of PARG was presumably due to the non-covalent interactions of these proteins with free ADP-ribose polymers. Thus, the presence of histone and nuclear matrix proteins should be taken into consideration when measuring PARG activity.
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评估固定化硼酸盐在腺嘌呤和吡啶核苷酸代谢研究中的应用。
影响因子：
2.9
作者：
Alvarez-Gonzalez,R;Juarez-Salinas,H;Jacobson,EL;Jacobson,MK
通讯作者：
Jacobson,MK
影响因子：
2.9
作者：
ALVAREZGONZALEZ, R;JACOBSON, MK
通讯作者：
JACOBSON, MK
DOI：
10.1016/s0074-7696(08)61228-0
发表时间：
1995
期刊：
International review of cytology
影响因子：
--
作者：
Ronald Berezney;M. Mortillaro;Hong Ma;Xiangyun Wei;J. Samarabandu
通讯作者：
Ronald Berezney;M. Mortillaro;Hong Ma;Xiangyun Wei;J. Samarabandu
ADP-核糖的内源性聚合物与核基质相关。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Cardenas-Corona,ME;Jacobson,EL;Jacobson,MK
通讯作者：
Jacobson,MK