Name: simultaneous measurement of hdac1 and hdac6 activity in hela cells using uhplc-ms
Uploaded: 2017-08-10T12:30:00
Description: Watch this Scientific Journal Video about Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS at JoVE.com

作者承认成本行动 CM1406 （表观遗传化学生物学）。研究报告在这份出版物由皮埃尔 · 米谢尔基金会支持。

1.细胞培养
注： 在标准组培罩执行以下步骤。不育技术熟悉预计。
<ol>
	<li>文化 80%融合在 T75 细胞培养瓶中补充含 10%胎牛血清，青霉素 G 的 MEM HeLa 细胞 (100 U/mL)，和链霉素 （100 毫克/毫升）。 注： 细胞培养、 治疗和裂解步骤在层流流动，与细胞孵育步骤在执行 5%CO2在无菌条件下湿大气中的 37 ° C 下进行。</li>
	<li>洗细胞两次将 5 毫升、 吸出新方案，和添加 1 毫?...

<ol>
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Study of ACY-1215 in combination with lenalidomide, and dexamethasone in multiple myeloma Available from: <a target="_blank" href="https://clinicaltrials.gov/ct2/show/NCT01583283">https://clinicaltrials.gov/ct2/show/NCT01583283</a> (2012)</li><li>. 13ACY-1215 (ricolinostat) in combination with pomalidomide and low-dose dex in relapsed-and-refractory multiple myeloma Available from: <a target="_blank" href="https://clinicaltrials.gov/ct2/show/NCT01997840">https://clinicaltrials.gov/ct2/show/NCT01997840</a> (2013)</li><li>. Study of ACY-241 alone and in combination with pomalidomide and dexamethasone in multiple myeloma Available from: <a target="_blank" href="https://clinicaltrials.gov/ct2/show/NCT02400242">https://clinicaltrials.gov/ct2/show/NCT02400242</a> (2015)</li><li>Yee, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ricolinostat+plus+lenalidomide,+and+dexamethasone+in+relapsed+or+refractory+multiple+myeloma:+a+multicentre+phase+1b+trial.">Ricolinostat plus lenalidomide, and dexamethasone in relapsed or refractory multiple myeloma: a multicentre phase 1b trial.</a> Lancet Oncol. 17 (11), 1569-1578 (2016).</li><li>Jung, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homogenous+non-isotopic+assays+for+histone+deacetylase+activity.">Homogenous non-isotopic assays for histone deacetylase activity.</a> Expert Opin Ther Pat. 13 (6), 935 (2003).</li><li>Zwick, V., Sim&#245;es-Pires, C., Cuendet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-based+multi-substrate+assay+coupled+to+UHPLC-ESI-MS/MS+for+a+quick+identification+of+class-specific+HDAC+inhibitors.">Cell-based multi-substrate assay coupled to UHPLC-ESI-MS/MS for a quick identification of class-specific HDAC inhibitors.</a> J Enzyme Inhibi Med Chem. 31 (1), 209-214 (2016).</li><li>Khan, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+the+class+and+isoform+selectivity+of+small-molecule+histone+deacetylase+inhibitors.">Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors.</a> Biochem J. 409 (2), 581-589 (2008).</li><li>Glaser, K. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+protein+acetylation+induced+by+novel+histone+deacetylase+inhibitors.">Differential protein acetylation induced by novel histone deacetylase inhibitors.</a> Biochem Biophys Res Commun. 325 (3), 683-690 (2004).</li><li>Butler, K. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rational+design+and+simple+chemistry+yield+a+superior,+neuroprotective+HDAC6+inhibitor,+tubastatin+A.">Rational design and simple chemistry yield a superior, neuroprotective HDAC6 inhibitor, tubastatin A.</a> J Am Chem Soc. 132 (31), 10842-10846 (2010).</li><li>Jung, M., Yong, K. -. J., Velena, A., Lee, S., Sippl, W., Jung, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-Based+Assays+for+HDAC+Inhibitor+Hit+Validation.">Cell-Based Assays for HDAC Inhibitor Hit Validation.</a> Epigenetic Targets in Drug Discovery. , (2010).</li><li>Milli, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomic+analysis+of+cellular+response+to+novel+proapoptotic+agents+related+to+atypical+retinoids+in+human+IGROV-1+ovarian+carcinoma+cells.">Proteomic analysis of cellular response to novel proapoptotic agents related to atypical retinoids in human IGROV-1 ovarian carcinoma cells.</a> J Proteome Res. 10 (3), 1191-1207 (2011).</li><li>Zwick, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+a+selective+HDAC6+inhibitor+active+in+neuroblasts.">Synthesis of a selective HDAC6 inhibitor active in neuroblasts.</a> Bioorg Med Chem Lett. 26 (20), 4955-4959 (2016).</li><li>Ciossek, T., Julius, H., Wieland, H., Maier, T., Beckers, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+homogeneous+cellular+histone+deacetylase+assay+suitable+for+compound+profiling+and+robotic+screening.">A homogeneous cellular histone deacetylase assay suitable for compound profiling and robotic screening.</a> Anal Biochem. 372 (1), 72-81 (2008).</li><li>Heltweg, B., Dequiedt, F., Marshall, B. 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HDAC 抑制是一个热门话题，药物发现，当前的重点放在为癌症治疗21HDAC6 选择性抑制剂。HDAC 选择性通常是由一系列的高吞吐量、 无细胞化验，打算对个别 HDAC 异构体21抑制效价评定的。然而，一种抑制剂的选择性必须另外确认在活细胞中通过评估内源性蛋白质底物，如组蛋白的乙酰化状态 （为 I 类乙酰） 和微管蛋白 （为 HDAC6)。因此，细胞化验涉及抗体识别 ?...

乙酰属于一个家庭能够脱乙酰度内的染色质结构的组蛋白的酶。他们还在胞浆内有其他蛋白质底物和位于中各单元格舱室。共 18 HDAC 异构体迄今为止识别出已涉及到几种细胞机制，包括转录因子基因的表达，以及细胞信号转导的调控和运输1，，23，4，5，，67。催化 HDAC 抑制剂已成为潜在的治疗药物，用于治疗癌症。大多数的 HDAC 抑制剂，目前被 FDA 批准为 T 细胞淋巴瘤和多发性骨髓瘤的治疗，非特定 HDAC 抑制剂，如 vorinostat (SAHA)、 belinostat 和 panobinostat8，9。然而，一系列的副作用已与泛-抑制剂和亚型特异性小分子搜索是一个热门话题，在药物化学、 新药开发。因此，类我 （HDAC1 3 和 HDAC8） 选择性抑制剂 romidepsin 是已经核准的药物10，而 HDAC6 特异性抑制剂，目前正在进行的临床实验，在多发性骨髓瘤11，，1213，，1415增加治疗潜力。
筛查检测来表征 HDAC 抑制剂都基于 HDAC 基板与酶源 （单一异构体、 核提取物或细胞裂解液） 的孵化。承印物上通常是含耦合到裂解荧光 （例如，香豆素），乙酰基赖氨酸残基的小分子多肽序列，如 N-(4-methyl-7-aminocoumarinyl)-Nα-(t-butoxycarbonyl)-Nω-acetyllysineamide (MAL)16。为了区分异构体的具体活动，涉及每个异构体的分离无细胞化验是必要和可能不会反映在活细胞中的真正异构体活动。异构体特定底物市面上，如苄基 (S)-[1-(4-methyl-2-oxo-2H-chromen-7-ylcarbamoyl)-5-propionylaminopentyl]carbamate (MOCPAC，HDAC1 具体基板) 和 (S)-[5-acetylamino-1-(2-oxo-4-trifluoromethyl-2H-chromen-7-ylcarbamoyl)pentyl]carbamic 酸叔丁酯 (BATCP，HDAC6 具体基板) (图 1B)。然而，仲裁法 》 的多基质混合物中，MOCPAC 和 BATCP 给活细胞将不允许个别的脱乙酰度产品检测荧光法测定，既然他们承担相同的裂解荧光。
这里介绍的方法可以检测和相对定量方法的每个衬底和 HeLa 细胞使用紧接着 UHPLC-质谱-质谱分析17多底物测定其脱乙酰基的产品。HDAC 检测被进行对 HeLa 细胞能够直接识别的 HDAC 抑制活性和特异性的测试化合物对内源性乙酰。那里是专注于 HDAC1 和 HDAC6，同时评估。为了实现在单一孵化测定这些酶法测量，非异性和特异性 HDAC 基质混合物被添加到在 96 孔板上镀的处理和未经处理 HeLa 细胞。后孵化的一步，细胞的裂解释放基质和他们各自的反应产物，这分离和检测 UHPLC 气相色谱-质谱法 (图 1)。仲裁法 》，MOCPAC 和 BATCP 的底物的脱乙酰度的产品分别是脱乙酰度的玛 (dMAL)、 脱乙酰度的 MOCPAC (dMOCPAC) 和脱乙酰度的 BATCP (dBATCP)。剂量-反应曲线可以用活性的化合物。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55878/55878fig1.jpg"> 
图 1: 基于单元格的 HDAC 法通过多个基板 UHPLC 气相色谱-质谱分析鉴定特定于 HDAC1 和 HDAC6 抑制剂的一般计划。(A) 计划的典型 96 孔板化合物治疗 （测试） 和 HeLa 细胞 （对照），以及无细胞的空白。(B) 补充说： 作为一种混合物 (每个 21 µ M)，由内源性乙酰脱乙酰基底的化学结构。(C) 典型 UHPLC-质谱联用色谱显示添加衬底 （仲裁法 》，MOCPAC 和 BATCP） 和脱乙酰度的产品的峰值 (dMAL，dMOCPAC，和 dBACTP，分别)。<a href="http://ecsource.jove.com/files/ftp_upload/55878/55878fig1large.jpg" target="_blank">请点击这里查看此图的大版本。</a>

<table border="0" cellpadding="0" cellspacing="0" width="887">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">BATCP</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">B4061</td>
			<td style="width:273px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MAL</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td>SCP0168</td>
			<td>Synonym: BOC-Ac-Lys-AMC&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">MOCPAC</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td>M2195</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">MS275</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">EPS002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Trichostatin A</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">T8552</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Tubastatin A</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">SML0044</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Acetonitrile, HPLC grade</td>
			<td style="width:191px;">Fisher Scientific</td>
			<td style="width:152px;">10660131</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Formic acid, LC/MS grade</td>
			<td style="width:191px;">Fisher Scientific</td>
			<td style="width:152px;">10596814</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:271px;">H2O, HPLC grade</td>
			<td style="width:191px;"></td>
			<td style="width:152px;"></td>
			<td>distilled H2O filtered through a Milli-Q purification system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">HeLa cells</td>
			<td style="width:191px;">ATCC</td>
			<td style="width:152px;">ATCC CRM-CCL-2</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cell dissociation reagent TrypLE Express</td>
			<td style="width:191px;">ThermoFisher Scientific</td>
			<td style="width:152px;">12604013</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">DMSO, cell culture grade</td>
			<td style="width:191px;">Applichem</td>
			<td style="width:152px;">146463</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">DPBS</td>
			<td style="width:191px;">ThermoFisher Scientific</td>
			<td>14190144</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Fetal bovine serum&#160;</td>
			<td style="width:191px;">Biowest</td>
			<td>S1810</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">MEM</td>
			<td style="width:191px;">ThermoFisher Scientific</td>
			<td>22561021</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Penicillin-Streptomycin</td>
			<td style="width:191px;">Bioconcept</td>
			<td>4-01F00-H&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10X RIPA buffer</td>
			<td style="width:191px;">Abcam</td>
			<td style="width:152px;">ab156034</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">SigmaFast protease inhibitor tablets</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">S8820</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">96-well plates, sterile, flat-bottom, tissue culture treated Corning</td>
			<td style="width:191px;">VWR</td>
			<td style="width:152px;">29442-058</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">96-well plates, non-sterile, V-bottom Corning</td>
			<td style="width:191px;">VWR</td>
			<td style="width:152px;">29442-404</td>
			<td>used in the centrifugation step</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">96-well plate, conical bottom, Nunc</td>
			<td style="width:191px;">ThermoFisher Scientific</td>
			<td style="width:152px;">249944</td>
			<td>compatible with the Acquity UHPLC system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">T75&#160; cell culture flasks Corning</td>
			<td style="width:191px;">Sigma-Aldrich</td>
			<td style="width:152px;">CLS430641&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Peelable heat sealing foil&#160;</td>
			<td style="width:191px;">Waters</td>
			<td style="width:152px;">186002789</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Acquity UPLC system</td>
			<td style="width:191px;">Waters</td>
			<td style="width:152px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Eppendorf Centrifuge 5810 R</td>
			<td style="width:191px;">Fisher Scientific</td>
			<td style="width:152px;">05-413-323</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Integration software: MassLynx V4.1</td>
			<td style="width:191px;">Waters</td>
			<td style="width:152px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:271px;">Combi thermo-sealer SP-0669/240</td>
			<td style="width:191px;">Waters</td>
			<td style="width:152px;"></td>
			<td>Catalog number not available</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:271px;">Quattro micro API Tandem Quadrupole System</td>
			<td style="width:191px;">Waters</td>
			<td style="width:152px;"></td>
			<td>Catalog number not available</td>
		</tr>
	</tbody>
</table>

simultaneous measurement of hdac1 and hdac6 activity in hela cells using uhplc-ms

寻找新的组蛋白去乙酰化酶 (HDAC) 抑制剂是提高药物发现感兴趣。异构体选择性以来，在聚光灯下的 romidepsin，一类批准我 HDAC 抑制剂用于癌症治疗和 HDAC6 特异性抑制剂对多发性骨髓瘤的临床研究。本方法用于确定测试化合物 HDAC1 和 HDAC6 细胞的抑制活性。使用超高性能液相色谱 — — 质谱 (UHPLC-MS) 分析方法对特定的底物与处理和未经处理的 HeLa 细胞培养，测定了异构体活性。该方法的优点是反映内的细胞环境，与无细胞的生化研究孤立异构体的内源性 HDAC 活动。此外，因为它基于定量化合成基质，方法不需要内源性乙酰化蛋白抗体识别。它很容易适用于几种细胞系和一个自动化的过程。该方法已经证明有用找到 HDAC6 选择性化合物在神经母细胞。代表性的结果如下所示的标准的 HDAC 抑制剂曲 （非特定），MS275 （特定的 HDAC1） 和 tubastatin （HDAC6-特定） 使用 HeLa 细胞。

能够识别中的应用方法非选择性和选择性抑制剂为 HDAC1 （第一类） 和 HDAC6 （类 IIb），HeLa 细胞治疗与已知的标准化合物： 曲古菌素 A （非选择性 HDAC1 和 HDAC6 之间）18、19MS275 （HDAC1 选择性抑制剂） 和 tubastatin （HDAC6 选择性抑制剂）20。使用本方法 (图 1)，IC50值可以直接确定在活细胞中 ...

Watch this Scientific Journal Video about Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS at JoVE.com

Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS

本方法用于确定亚型特异性抑制剂组蛋白去乙酰化酶 (HDAC) 在 HeLa 细胞中由多个基板 UHPLC 气相色谱-质谱分析。这是一种抗体免费的方法，开发以反映生活的 HDAC1 和 HDAC6 活性与单一异构体无细胞检测的细胞环境。

同时测量 HDAC1 和 HDAC6 活动在 HeLa 细胞使用 UHPLC 女士

The search for new histone deacetylase (HDAC) inhibitors is of increasing interest in drug discovery. Isoform selectivity has been in the spotlight since ...

Research

JoVE Journal

Chemistry

Steady-state, Pre-steady-state, and Single-turnover Kinetic Measurement for DNA Glycosylase Activity

稳态，稳定前的状态，单营业额动力学测量DNA糖基化酶活性

Human 8-oxoguanine DNA glycosylase (OGG1) excises the mutagenic oxidative DNA lesion 8-oxo-7,8-dihydroguanine (8-oxoG) from DNA. Kinetic characterization ...

科研

化学

A Simple and Rapid Protocol for Measuring Neutral Lipids in Algal Cells Using Fluorescence

一种简单快速的协议用荧光测量中性脂肪在藻细胞

Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal ...

HPLC Measurement of the DNA Oxidation Biomarker, 8-oxo-7,8-dihydro-2&#8217;-deoxyguanosine, in Cultured Cells and Animal Tissues

HPLC Measurement of the DNA Oxidation Biomarker, 8-oxo-7,8-dihydro-2ÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢-deoxyguanosine, in Cultured Cells and Animal Tissues

该DNA氧化标志物的高效液相色谱法测量，8-氧代-7,8-二氢-2&#39;-脱氧鸟苷，在培养细胞和动物组织

Oxidative stress is associated with many physiological and pathological processes, as well as xenobiotic metabolism, leading to the oxidation of ...

Dynamic Electrochemical Measurement of Chloride Ions

氯离子的动态电化学测试

This protocol describes the dynamic measurement of chloride ions using the transition time of a silver silver chloride (Ag/AgCl) electrode. Silver silver ...

Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical (Methylation) and Physical (Mass Spectrometry, Nuclear Magnetic Resonance) Techniques

Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical Methylation and Physical Mass Spectrometry, Nuclear Magnetic Resonance Techniques

使用酶法，化学（甲基化）和物理（质谱，核磁共振）技术，植物墙Heteroxylans测序

This protocol describes the specific techniques used for the characterization of reducing end (RE) and internal region glycosyl sequence(s) of ...

Fabrication and Testing of Photonic Thermometers

光子温度计的制作与测试

In recent years, a push for developing novel silicon photonic devices for telecommunications has generated a vast knowledge base that is now being ...

Disentangling High Strength Copolymer Aramid Fibers to Enable the Determination of Their Mechanical Properties

解析高强共聚物芳纶纤维使其力学性能的测定

Traditionally, soft body armor has been made from poly(p-phenylene terephthalamide) (PPTA) and ultra-high molecular weight polyethylene. However, to ...

High Resolution Physical Characterization of Single Metallic Nanoparticles

单金属纳米粒子的高分辨率物理特性

Individual molecules can be detected and characterized by measuring the degree by which they reduce the ionic current flowing through a single ...

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy

A New Straightforward Method for Lipophilicity logP Measurement using 19F NMR Spectroscopy

一种利用19f 核磁共振光谱测定脂质性 (日志 p) 的新方法

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction ...

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

基于NMR的活性测定,用于测定核苷裂解酶的化合物抑制、IC50值、伪影活性和全细胞活性

NMR spectroscopy is often used for the identification and characterization of enzyme inhibitors in drug discovery, particularly in the context of fragment ...