H-Ala-Pro-pNA*HCl | 65022-17-5

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 羧酸及其衍生物
• 英文名称: H-Ala-Pro-pNA*HCl
• 英文别名: H-Ala-Pro-pNA HCl；(2S)-1-[(2S)-2-aminopropanoyl]-N-(4-nitrophenyl)pyrrolidine-2-carboxamide;hydrochloride
• CAS: 65022-17-5
• 化学式: C₁₄H₁₈N₄O₄*ClH
• mdl: MFCD00155539
• 分子量: 342.782
• InChiKey: MYWOEAKMMDWWIZ-CSDGMEMJSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  0.53
• 重原子数：
  23
• 可旋转键数：
  3
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.428
• 拓扑面积：
  121
• 氢给体数：
  3
• 氢受体数：
  5

反应信息

• 作为反应物：
  描述：

  叔丁氧羰基-L-天冬氨酸-4-叔丁酯 、 H-Ala-Pro-pNA*HCl 在 N-甲基吗啉 、 氯甲酸异丁酯 、 盐酸 作用下， 以 四氢呋喃 、 1,4-二氧六环 、 水 为溶剂， 反应 2.33h， 生成
  参考文献：
  名称：

  Continuous assays for meprin alpha and beta using prolyl tripeptidyl aminopeptidase (PtP) from Porphyromonas gingivalis

  摘要：

  Common assays for endoprotease activity of meprin alpha and beta are based on cleavage of internally quenched substrates. Although direct and convenient, for meprins these assays bear disadvantages such as, e.g., significant substrate inhibition or potential fluorescence quenching by compounds applied in inhibitor analysis. Here, we present a novel continuous assay by introducing an auxiliary enzyme, prolyl tripeptidyl aminopeptidase (PtP) and the chromogenic substrate KKGYVADAP-p-nitroanilide. We provide a quick strategy for expression and one-step-purification of the auxiliary enzyme. The enzyme kinetic data for meprin alpha and beta suggest hyperbolic v/Scharacteristics, the kinetic parameters of substrate conversion by meprin beta were K-m = 184 +/- 32 mu M and k(cat) = 20 +/- 4 s(-1). We also present conditions for the use of the fluorogenic substrate KKGYVADAP-AMC to assess meprin beta activity. The assays were applied for determination of inhibitory parameters of the natural inhibitor actinonin and two recently published hydroxamates. Hence, we present two novel methods, which can be applied to assess inhibitory mechanism and potency with the attractive current drug targets meprin alpha and beta. Furthermore, the assay might also provide implications for analysis of other endoproteases as well as their inhibitors.

  DOI：

  10.1016/j.ab.2018.08.005
• 作为产物：
  描述：

  N-叔丁氧羰基-L-丙氨酸 、 (S)-N-(4-nitrophenyl)pyrrolidine-2-carboxamide hydrochloride 在 N-甲基吗啉 、 氯甲酸异丁酯 、 盐酸 作用下， 以 四氢呋喃 、 1,4-二氧六环 、 水 为溶剂， 反应 2.33h， 生成 H-Ala-Pro-pNA*HCl
  参考文献：
  名称：

  Continuous assays for meprin alpha and beta using prolyl tripeptidyl aminopeptidase (PtP) from Porphyromonas gingivalis

  摘要：

  Common assays for endoprotease activity of meprin alpha and beta are based on cleavage of internally quenched substrates. Although direct and convenient, for meprins these assays bear disadvantages such as, e.g., significant substrate inhibition or potential fluorescence quenching by compounds applied in inhibitor analysis. Here, we present a novel continuous assay by introducing an auxiliary enzyme, prolyl tripeptidyl aminopeptidase (PtP) and the chromogenic substrate KKGYVADAP-p-nitroanilide. We provide a quick strategy for expression and one-step-purification of the auxiliary enzyme. The enzyme kinetic data for meprin alpha and beta suggest hyperbolic v/Scharacteristics, the kinetic parameters of substrate conversion by meprin beta were K-m = 184 +/- 32 mu M and k(cat) = 20 +/- 4 s(-1). We also present conditions for the use of the fluorogenic substrate KKGYVADAP-AMC to assess meprin beta activity. The assays were applied for determination of inhibitory parameters of the natural inhibitor actinonin and two recently published hydroxamates. Hence, we present two novel methods, which can be applied to assess inhibitory mechanism and potency with the attractive current drug targets meprin alpha and beta. Furthermore, the assay might also provide implications for analysis of other endoproteases as well as their inhibitors.

  DOI：

  10.1016/j.ab.2018.08.005

文献信息

• Continuous assays for meprin alpha and beta using prolyl tripeptidyl aminopeptidase (PtP) from Porphyromonas gingivalis
  作者：Anja Schulze、Michael Wermann、Hans-Ulrich Demuth、Tadashi Yoshimoto、Daniel Ramsbeck、Dagmar Schlenzig、Stephan Schilling

  DOI：10.1016/j.ab.2018.08.005

  日期：2018.10

  Common assays for endoprotease activity of meprin alpha and beta are based on cleavage of internally quenched substrates. Although direct and convenient, for meprins these assays bear disadvantages such as, e.g., significant substrate inhibition or potential fluorescence quenching by compounds applied in inhibitor analysis. Here, we present a novel continuous assay by introducing an auxiliary enzyme, prolyl tripeptidyl aminopeptidase (PtP) and the chromogenic substrate KKGYVADAP-p-nitroanilide. We provide a quick strategy for expression and one-step-purification of the auxiliary enzyme. The enzyme kinetic data for meprin alpha and beta suggest hyperbolic v/Scharacteristics, the kinetic parameters of substrate conversion by meprin beta were K-m = 184 +/- 32 mu M and k(cat) = 20 +/- 4 s(-1). We also present conditions for the use of the fluorogenic substrate KKGYVADAP-AMC to assess meprin beta activity. The assays were applied for determination of inhibitory parameters of the natural inhibitor actinonin and two recently published hydroxamates. Hence, we present two novel methods, which can be applied to assess inhibitory mechanism and potency with the attractive current drug targets meprin alpha and beta. Furthermore, the assay might also provide implications for analysis of other endoproteases as well as their inhibitors.