Suc-丙氨酰-丙氨酰-丙氨酰-对硝基苯胺 | 52299-14-6

• 物质功能分类: 生物化工 - 氨基酸及其衍生物 - 丙氨酸类衍生物
• 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
• 中文名称: Suc-丙氨酰-丙氨酰-丙氨酰-对硝基苯胺
• 中文别名: N-琥珀酰-丙氨酸-丙氨酸-丙氨酸-对硝基苯胺；N-琥珀酰-L-丙氨酰-L-丙氨酰-L-丙氨酸
• 英文名称: N-succinyl-Ala-Ala-Ala-p-nitroanilide
• 英文别名: Suc-Ala-Ala-Ala-pNA；N-succinyl-L-alanyl-L-alanyl-L-alanine 4-nitroanilide；N-suc-(Ala)-Ala-Ala-nitroanilide；N-Suc-Ala-Ala-Ala-p-nitroanilide；SANA；Succinyl-trialanine-4-nitroanilide；4-[[(2S)-1-[[(2S)-1-[[(2S)-1-(4-nitroanilino)-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-4-oxobutanoic acid
• CAS: 52299-14-6
• 化学式: C₁₉H₂₅N₅O₈
• 分子量: 451.436
• InChiKey: GVUGADOWXGKRAE-SRVKXCTJSA-N

物化性质

• 沸点：
  923.5±65.0 °C(Predicted)
• 密度：
  1.370±0.06 g/cm3(Predicted)
• 溶解度：
  DMF：3mg/mL； DMSO：5mg/mL；乙醇：微溶； PBS（pH 7.2）：0.3 mg/mL
• 稳定性/保质期：

  如果按照规格使用和储存，则不会分解，未有已知危险反应。

计算性质

• 辛醇/水分配系数(LogP)：
  -0.4
• 重原子数：
  32
• 可旋转键数：
  10
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.42
• 拓扑面积：
  200
• 氢给体数：
  5
• 氢受体数：
  8

安全信息

• 安全说明：
  S24/25
• WGK Germany：
  3
• 海关编码：
  29242990
• 危险性防范说明：
  P264,P280,P302+P352,P337+P313,P305+P351+P338,P362+P364,P332+P313
• 危险性描述：
  H315,H319

反应信息

• 作为反应物：
  描述：

  Suc-丙氨酰-丙氨酰-丙氨酰-对硝基苯胺 以 水 为溶剂， 反应 10.0h， 生成 4-硝基苯胺
  参考文献：
  名称：

  Stereoisomeric alanine peptides as substrates for human spleen fibrinolytic proteinase (SFP).

  摘要：

  与猪胰弹性蛋白酶相比，本研究测试了作为人脾纤维蛋白溶解酶（SFP）底物的四种立体异构体 Z-Ala-Ala-OMe 和八种立体异构体 Z-Ala-Ala-Ala-OMe。这两种酶不仅对 Z-L-Ala-L-Ala-L-Ala-Ome（VI），而且对 Z-D-Ala-L-Ala-L-Ala-Ome（VII）都表现出酯酶活性。虽然弹性蛋白酶对 Suc-L-Ala-L-Ala-L-Ala-pNA （XVI）的酰胺分解速度是 SFP 的十倍，但弹性蛋白酶对 VI 的酯解速度仅是 SFP 的两倍左右。

  DOI：

  10.1248/cpb.29.1762
• 作为产物：
  描述：

  benzyl ((S)-1-(((S)-1-hydrazinyl-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate 在 盐酸 、 Zeolit G (OH form) 、 氢溴酸 、 sodium nitrite 作用下， 以 溶剂黄146 、 N,N-二甲基甲酰胺 为溶剂， 反应 1.0h， 生成 Suc-丙氨酰-丙氨酰-丙氨酰-对硝基苯胺
  参考文献：
  名称：

  Kasafirek, Evzen; Fric, Premysl; Slaby, Jan, Collection of Czechoslovak Chemical Communications, 1987, vol. 52, # 6, p. 1625 - 1633

  摘要：

  DOI：

文献信息

• Specificity of a membrane-bound neutral endopeptidase from rat kidney.
  作者：YOSHIMITSU SHIMAMORI、YASUHIRO WATANABE、YUKIO FUJIMOTO

  DOI：10.1248/cpb.34.275

  日期：——

  The substrate specificity of a purified kidney neutral endopeptidase was studied. The endopeptidase hydrolyzed a variety of biologically active peptides, such as angiotensins (angiotensins I, II and III), bradykinins (bradykinin, Lys-bradykinin, Met-Lys-bradykinin and des-9Arg-bradykinin), enkephalins (Leu-enkephalin and Met-enkephalin), neurotensin and substance P, and was found to cleave only the bonds at the amino side of hydrophobic amino acids in the peptides. However, when a hydrophobic amino acid was present at the C-terminus or at the position adjacent to the N-terminus, the bond of the hydrophobic residue was not cleaved. In further studies on the degradation of a series of homo-oligopeptides, the enzyme appeared to hydrolyze those consisting of at least a tetrapeptide unit of hydrophobic amino acids such as Ala and Phe. The specificity of the membrane-bound neutral endopeptidase from rat kidney indicated by these results can be summarized as follows : the enzyme requires at least a tetrapeptide unit for hydrolysis, and cleavage occurs only at the amino side of a hydrophobic amino acid, when one is present at the third position of the tetrapeptide unit.

  纯化的肾脏中性内切肽酶的底物特异性被研究。该内切肽酶水解多种生物活性肽，如血管紧张素（血管紧张素I、II和III）、舒缓激肽（舒缓激肽、赖氨酸-舒缓激肽、甲硫氨酸-赖氨酸-舒缓激肽和去-9精氨酸-舒缓激肽）、脑啡肽（亮氨酸-脑啡肽和甲硫氨酸-脑啡肽）、神经降压素和P物质，并且发现它仅在肽中疏水氨基酸的氨基侧切割肽键。然而，当疏水氨基酸位于C末端或紧邻N末端的位置时，疏水残基的肽键不会被切割。在进一步研究一系列同源寡肽的降解过程中，该酶似乎水解那些至少由疏水氨基酸如丙氨酸和苯丙氨酸组成四肽单位的肽。根据这些结果，大鼠肾脏膜结合中性内切肽酶的特异性可以总结如下：酶需要至少一个四肽单位进行水解，并且仅在疏水氨基酸存在于四肽单位的第三位置时，才在该疏水氨基酸的氨基侧进行切割。
• The molecular basis of the effect of temperature on enzyme activity
  作者：Roy M. Daniel、Michelle E. Peterson、Michael J. Danson、Nicholas C. Price、Sharon M. Kelly、Colin R. Monk、Cristina S. Weinberg、Matthew L. Oudshoorn、Charles K. Lee

  DOI：10.1042/bj20091254

  日期：2010.1.15

  Experimental data show that the effect of temperature on enzymes cannot be adequately explained in terms of a two-state model based on increases in activity and denaturation. The Equilibrium Model provides a quantitative explanation of enzyme thermal behaviour under reaction conditions by introducing an inactive (but not denatured) intermediate in rapid equilibrium with the active form. The temperature midpoint (Teq) of the rapid equilibration between the two forms is related to the growth temperature of the organism, and the enthalpy of the equilibrium (ΔHeq) to its ability to function over various temperature ranges. In the present study, we show that the difference between the active and inactive forms is at the enzyme active site. The results reveal an apparently universal mechanism, independent of enzyme reaction or structure, based at or near the active site, by which enzymes lose activity as temperature rises, as opposed to denaturation which is global. Results show that activity losses below Teq may lead to significant errors in the determination of ΔG*cat made on the basis of the two-state (‘Classical’) model, and the measured kcat will then not be a true indication of an enzyme's catalytic power. Overall, the results provide a molecular rationale for observations that the active site tends to be more flexible than the enzyme as a whole, and that activity losses precede denaturation, and provide a general explanation in molecular terms for the effect of temperature on enzyme activity.

  实验数据表明，温度对酶的影响无法用基于活性增加和变性的双态模型来充分解释。平衡模型通过引入与活性形式快速平衡的非活性（但未变性）中间体，对反应条件下的酶热行为提供了定量解释。两种形式之间快速平衡的温度中点（Teq）与生物体的生长温度有关，而平衡焓（ΔHeq）与生物体在不同温度范围内发挥作用的能力有关。在本研究中，我们发现活性和非活性形式之间的区别在于酶的活性位点。研究结果揭示了一种与酶的反应或结构无关的、基于活性位点或活性位点附近的明显普遍机制，通过这种机制，酶的活性随着温度的升高而丧失，而变性则是全球性的。研究结果表明，低于 Teq 的活性损失可能会导致根据双态（"经典"）模型确定的 ΔG*cat 出现重大误差，因此测得的 kcat 无法真实反映酶的催化能力。总之，这些结果为观察到的活性位点往往比酶整体更灵活、变性前会出现活性损失等现象提供了分子原理，并从分子角度为温度对酶活性的影响提供了一般性解释。
• A succinyl-trialanine p-nitroanilide hydrolase in hog kidney cytosol: Its identification as proline endopeptidase.
  作者：SHINJI SOEDA、MASANORI OHYAMA、ATSUO NAGAMATSU

  DOI：10.1248/cpb.32.1510

  日期：——

  A succinyl-trialanine p-nitroanilide [Suc-(Ala)3-pNA] hydrolase which is able to hydrolyze an artificial elastase substrate, Suc-(Ala)3-pNA, but unable to hydrolyze a naturally occurring substrate, elastin, was highly purified from hog kidney cytosol. The apparent molecular weight of the enzyme was estimated to be 65000 by gel filtration on Sephadex G-150 and 68000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the isoelectric point of the enzyme was 5.0. The enzyme is an endopeptidase which catalyzes the hydrolysis of peptides with the general structure Y-Ala (or Pro)-X (Y=peptide or N-protected amino acid ; X=amino acid moiety, peptide or amide) at the carboxyl side of alanine and proline residues. The enzyme was markedly inhibited by diisopropyl fluorophosphate and p-chloromercuribenzoate. Ethylenediaminetetraacetate and 1, 10-phenanthroline, however, were not inhibitors of the enzyme. The enzyme activity was retained on an affinity column having a proline endopeptidase [EC 3.4.21.26] inhibitor, Z-Gly-Pro, as ligand and could be eluted at 0.125M NaCl (mean value). Suc-(Ala)3-pNA-hydrolytic activity coincided with the peak of proline endopeptidase activity as determined with a sensitive fluorogenic substrate, succinylglycyl-L-proline 4-methylcoumaryl-7-amide (Suc-Gly-Pro-MCA). The optimum pH and kcat/Km values (mM-1·s-1) were pH 7.5 and 5.4 for Suc-(Ala)3-pNA and pH 6.8 and 24.8 for Suc-Gly-Pro-MCA, and the enzyme activity was competitively inhibited by Z-Ala-Ala and Z-Gly-Pro, as is the case with proline endopeptidase. These results suggest that Suc-(Ala)3-pNA hydrolase in hog kidney cytosol may be identical with proline endopeptidase which was first found in human uterus as an oxytocindegrading enzyme.

  从猪肾细胞液中高度纯化了一种琥珀酰-鸟嘌呤对硝基苯胺[Suc-(Ala)3-pNA]水解酶，该酶能够水解人工弹性蛋白酶底物 Suc-(Ala)3-pNA，但不能水解天然底物弹性蛋白。通过在 Sephadex G-150 上进行凝胶过滤，估计该酶的表观分子量为 65000，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，估计该酶的表观分子量为 68000，等电点为 5.0。该酶是一种内肽酶，可催化水解一般结构为 Y-Ala（或 Pro）-X（Y=肽或 N-保护氨基酸；X=氨基酸分子、肽或酰胺）的肽，其结构位于丙氨酸和脯氨酸残基的羧基侧。氟磷酸二异丙酯和对氯脲苯甲酸酯对该酶有明显的抑制作用。然而，乙二胺四乙酸盐和 1，10-菲罗啉不是该酶的抑制剂。以脯氨酸内肽酶[EC 3.4.21.26] 抑制剂 Z-Gly-Pro 为配体的亲和柱可保留酶的活性，并可在 0.125M NaCl（平均值）条件下洗脱。蔗糖-(Ala)3-pNA-水解活性与脯氨酸内肽酶活性的峰值相吻合，脯氨酸内肽酶活性是用敏感的荧光底物琥珀酰甘氨酰-L-脯氨酸 4-甲基香豆素-7-酰胺（Suc-Gly-Pro-MCA）测定的。Suc-(Ala)3-pNA 的最适 pH 值和 kcat/Km 值（mM-1-s-1）分别为 pH 7.5 和 5.4，Suc-Gly-Pro-MCA 的最适 pH 值和 kcat/Km 值（mM-1-s-1）分别为 pH 6.8 和 24.8。这些结果表明，猪肾细胞质中的 Suc-(Ala)3-pNA 水解酶可能与脯氨酸内肽酶相同。
• Substrate Specificity of Honeydew Melon Protease D, a Plant Serine Endopeptidase
  作者：Hiroo Yonezawa、Tetsuya Uchikoba、Makoto Kaneda

  DOI：10.1271/bbb.61.1277

  日期：1997.1

  The substrate specificity of honeydew melon (Cucumis melo var. inodorus Naud) protease D was studied by the use of synthetic substrates and oligopeptides derived from a protein hydrolyzate. The hydrolysis rates of succinyl-(L-Ala)1-3-p-nitroanilide (Suc-(Ala)1-3-pNA) the hydrolysis rate progressively rose in proportion to the increased chain length. Benzyloxycarbonyl-L-tyrosine p-nitrophenyl ester (Z-Tyr-ONp)

  通过使用合成底物和衍生自蛋白质水解产物的寡肽，研究了蜜瓜（Cucumis melo var。inodorus Naud）蛋白酶D的底物特异性。琥珀酰基-（L-Ala）1-3-对硝基苯胺（Suc-（Ala）1-3-pNA）的水解速率与链长的增加成比例地逐渐提高。蜜瓜蛋白酶D裂解了苄氧羰基-L-酪氨酸对硝基苯酯（Z-Tyr-ONp）和苯甲酰基-L-酪氨酸乙酯（Bz-Tyr-OEt），但苯甲酰-L-精氨酸对硝基苯胺（Bz -Arg-pNA），苄氧羰基-L-赖氨酸对硝基苯酯（Z-Lys-ONp）和甲苯磺酰基-L-精氨酸甲酯（Tos-Arg-OMe）未水解。与使用合成底物获得的结果相反，带电荷的氨基酸残基的羧基侧优先被寡肽底物中的酶裂解。在P2位置带电荷或极性氨基酸的底物没有被切割。另一方面，在P2处的非极性氨基酸或脯氨酸优选用于水解。获得了有关蛋白酶D亚位点的信息，可用于合成良好的底物。因为它与分
• Substrate Specificity of Aqualysin I, a Bacterial Thermophilic Alkaline Serine Protease from<i>Thermus aquaticus</i>YT-1: Comparison with Proteinase K, Subtilisin BPN′ and Subtilisin Carlsberg
  作者：Terumichi TANAKA、Hiroshi MATSUZAWA、Takahisa OHTA

  DOI：10.1271/bbb.62.2161

  日期：1998.1

  Aqualysin I is the alkaline serine protease isolated from an extreme thermophile, Thermus aquaticus YT-1. We analyzed kinetic properties of aqualysin I, using sixteen kinds of chromogenic succinyl-tripeptide p-nitroanilides as substrates. And we compared the substrate specificity of aqualysin I with those of proteinase K, subtilisin BPN′, and subtilisin Carlsberg. We found that aqualysin I had three subsites, S1, S2, and S3, in the substrate binding site. S1 site preferred alanine and phenylalanine. S2 site preferred alanine and norleucine. And S3 site preferred phenylalanine and isoleucine. These specificities were similar to those of proteinase K and subtilisin BPN′. The specificity of subtilisin Carlsberg differed from those of other enzymes.

  Aqualysin I 是从极端嗜热菌栖热菌 YT-1 中分离出来的碱性丝氨酸蛋白酶。我们以 16 种显色琥珀酰三肽对硝基苯胺为底物，分析了 aqualysin I 的动力学性质。我们还比较了 aqualysin I 与蛋白酶 K、枯草杆菌蛋白酶 BPN' 和枯草杆菌蛋白酶 Carlsberg 的底物特异性。我们发现 aqualysin I 在底物结合位点上有 3 个亚位点：S1、S2 和 S3。 S1位点优选丙氨酸和苯丙氨酸。 S2位点优选丙氨酸和正亮氨酸。而S3位点优选苯丙氨酸和异亮氨酸。这些特异性与蛋白酶 K 和枯草杆菌蛋白酶 BPN' 相似。嘉士伯枯草杆菌蛋白酶的特异性不同于其他酶。