Gly-Phe-MCA | 201852-70-2

• 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: Gly-Phe-MCA
• 英文别名: GF-MCA；L-Phenylalaninamide, glycyl-N-(4-methyl-2-oxo-2H-1-benzopyran-7-yl)-；(2S)-2-[(2-aminoacetyl)amino]-N-(4-methyl-2-oxochromen-7-yl)-3-phenylpropanamide
• CAS: 201852-70-2
• 化学式: C₂₁H₂₁N₃O₄
• 分子量: 379.415
• InChiKey: OXYDLMUFQZNZMD-KRWDZBQOSA-N

物化性质

• 沸点：
  723.8±60.0 °C(Predicted)
• 密度：
  1.315±0.06 g/cm3(Predicted)

计算性质

• 辛醇/水分配系数(LogP)：
  1.4
• 重原子数：
  28
• 可旋转键数：
  6
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.19
• 拓扑面积：
  111
• 氢给体数：
  3
• 氢受体数：
  5

反应信息

• 作为反应物：
  描述：

  Gly-Phe-MCA 在 乙二胺四乙酸 、 dipeptydyl-peptidase DPP₅ 、 sodium chloride 作用下， 以 aq. phosphate buffer 为溶剂， 反应 0.5h， 生成 甘氨酰苯基丙氨酸
  参考文献：
  名称：

  Identification and Characterization of Prokaryotic Dipeptidyl-peptidase 5 from Porphyromonas gingivalis

  摘要：

  Background: Dipeptidyl-peptidases (DPPs) are key factors for amino acid metabolism and bacterial growth of asaccharolytic Porphyromonas gingivalis. Results: DPP5, which is specific for Ala and hydrophobic residues, is expressed in the periplasmic space of P. gingivalis.Conclusion: DPP5 was discovered in prokaryotes for the first time. Significance: The discovery of DPP5 expands understanding of amino acid and energy metabolism in prokaryotes. Porphyromonas gingivalis, a Gram-negative asaccharolytic anaerobe, is a major causative organism of chronic periodontitis. Because the bacterium utilizes amino acids as energy and carbon sources and incorporates them mainly as dipeptides, a wide variety of dipeptide production processes mediated by dipeptidyl-peptidases (DPPs) should be beneficial for the organism. In the present study, we identified the fourth P. gingivalis enzyme, DPP5. In a dpp4-7-11-disrupted P. gingivalis ATCC 33277, a DPP7-like activity still remained. PGN_0756 possessed an activity indistinguishable from that of the mutant, and was identified as a bacterial orthologue of fungal DPP5, because of its substrate specificity and 28.5% amino acid sequence identity with an Aspergillus fumigatus entity. P. gingivalis DPP5 was composed of 684 amino acids with a molecular mass of 77,453, and existed as a dimer while migrating at 66 kDa on SDS-PAGE. It preferred Ala and hydrophobic residues, had no activity toward Pro at the P1 position, and no preference for hydrophobic P2 residues, showed an optimal pH of 6.7 in the presence of NaCl, demonstrated K-m and k(cat)/K-m values for Lys-Ala-MCA of 688 m and 11.02 m(-1) s(-1), respectively, and was localized in the periplasm. DPP5 elaborately complemented DPP7 in liberation of dipeptides with hydrophobic P1 residues. Examinations of DPP- and gingipain gene-disrupted mutants indicated that DPP4, DPP5, DPP7, and DPP11 together with Arg- and Lys-gingipains cooperatively liberate most dipeptides from nutrient oligopeptides. This is the first study to report that DPP5 is expressed not only in eukaryotes, but also widely distributed in bacteria and archaea.

  DOI：

  10.1074/jbc.m113.527333
• 作为产物：
  描述：

  L-phe-7-氨基-4-甲基香豆素 在 O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate 、 N,N-二异丙基乙胺 、 三氟乙酸 作用下， 以 二氯甲烷 、 乙腈 为溶剂， 生成 Gly-Phe-MCA
  参考文献：
  名称：

  人类组织蛋白酶C的第一个内部淬灭荧光底物的开发：在生物样品中酶的检测中的应用。

  摘要：

  组织蛋白酶C是一种广泛表达的半胱氨酸外肽酶，主要用于嗜中性粒细胞，细胞毒性T淋巴细胞和肥大细胞中与颗粒相关的促炎性丝氨酸蛋白酶的活化。已经显示出该酶可以在细胞外分泌。然而，其在人体液/生理样品中的发生还没有被彻底研究。在本研究过程中，设计并合成了用于测量人组织蛋白酶C活性的首个荧光共振能量转移肽。两个系列的四肽和五肽底物可以对组织蛋白酶C进行详细的S特异性研究，这是首次进行了研究。对获得的化合物进行的广泛酶研究导致选择了高度特异性和选择性的底物Thi-Ala（Mca）-Ser-Gly-Tyr（3-NO2）-NH2，该酶已成功用于组织蛋白酶C活性的检测在复杂的生物样品中，例如细胞裂解液，尿液和支气管肺泡灌洗液。进行所选底物的分子对接是为了更好地了解组织蛋白酶C活性位点中底物的结合模式。

  DOI：

  10.1016/j.abb.2016.10.007

文献信息

• A Judgment on Postmortem Aging in<i>Longissimus Dorsi</i>Based on a Peptide Substrate Library
  作者：Eiichiro ONITSUKA、Tomoyuki OKUMURA、Hiroshi MURAKAMI、Norikazu NISHINO、Fumiki MORIMATSU

  DOI：10.1271/bbb.60208

  日期：2006.12.23

  We attempted to develop a method to determine easily and effectively the degree of postmortem aging of pork longissimus dorsi (LD) by measuring the activity of proteases in the LD using fluorogenic peptide substrates. LD was used to measure the change with time in the protease activity detected with these substrates. Determining the variations within the LD muscles, strong positive correlations were found between changes in hardness and fluorescence intensities against Ac-Ala-MCA, Ac-Met-MCA, Ac-Ser-MCA, Ac-Thr-MCA, and Ac-Ala-Phe-MCA (P<0.005), and strong negative correlations were found between changes in total amounts of free amino acids and Ac-Ala-MCA, Ac-Met-MCA, Ac-Ser-MCA, Ac-Thr-MCA, and Ac-Ala-Phe-MCA (P<0.001). Negative correlations were also observed between changes in the amounts of free Ala, Arg, Lys, Leu, Met, Phe, and Tyr and the fluorescence intensities against Ala, Arg, Lys, Leu, Met, Phe, and Tyr-MCA respectively (P<0.001).

  我们尝试开发一种方法，通过使用荧光底物检测猪背最长肌（LD）中的蛋白酶活性，来简易有效地确定猪背最长肌死后老化的程度。利用这些底物检测到的蛋白酶活性的变化，用于衡量LD肌肉随时间的变化。在确定LD肌肉内的变异时，我们发现硬度的变化与荧光强度之间存在强烈的正相关，这些荧光强度是针对Ac-Ala-MCA、Ac-Met-MCA、Ac-Ser-MCA、Ac-Thr-MCA和Ac-Ala-Phe-MCA的（P<0.005），同时也发现总游离氨基酸量的变化与Ac-Ala-MCA、Ac-Met-MCA、Ac-Ser-MCA、Ac-Thr-MCA和Ac-Ala-Phe-MCA之间存在强烈的负相关（P<0.001）。我们还观察到，游离的Ala、Arg、Lys、Leu、Met、Phe和Tyr的量的变化与针对Ala、Arg、Lys、Leu、Met、Phe和Tyr-MCA的荧光强度之间存在负相关（P<0.001）。
• Identification and Characterization of Prokaryotic Dipeptidyl-peptidase 5 from Porphyromonas gingivalis
  作者：Yuko Ohara-Nemoto、Shakh M.A. Rouf、Mariko Naito、Amie Yanase、Fumi Tetsuo、Toshio Ono、Takeshi Kobayakawa、Yu Shimoyama、Shigenobu Kimura、Koji Nakayama、Keitarou Saiki、Kiyoshi Konishi、Takayuki K. Nemoto

  DOI：10.1074/jbc.m113.527333

  日期：2014.2

  Background: Dipeptidyl-peptidases (DPPs) are key factors for amino acid metabolism and bacterial growth of asaccharolytic Porphyromonas gingivalis. Results: DPP5, which is specific for Ala and hydrophobic residues, is expressed in the periplasmic space of P. gingivalis.Conclusion: DPP5 was discovered in prokaryotes for the first time. Significance: The discovery of DPP5 expands understanding of amino acid and energy metabolism in prokaryotes. Porphyromonas gingivalis, a Gram-negative asaccharolytic anaerobe, is a major causative organism of chronic periodontitis. Because the bacterium utilizes amino acids as energy and carbon sources and incorporates them mainly as dipeptides, a wide variety of dipeptide production processes mediated by dipeptidyl-peptidases (DPPs) should be beneficial for the organism. In the present study, we identified the fourth P. gingivalis enzyme, DPP5. In a dpp4-7-11-disrupted P. gingivalis ATCC 33277, a DPP7-like activity still remained. PGN_0756 possessed an activity indistinguishable from that of the mutant, and was identified as a bacterial orthologue of fungal DPP5, because of its substrate specificity and 28.5% amino acid sequence identity with an Aspergillus fumigatus entity. P. gingivalis DPP5 was composed of 684 amino acids with a molecular mass of 77,453, and existed as a dimer while migrating at 66 kDa on SDS-PAGE. It preferred Ala and hydrophobic residues, had no activity toward Pro at the P1 position, and no preference for hydrophobic P2 residues, showed an optimal pH of 6.7 in the presence of NaCl, demonstrated K-m and k(cat)/K-m values for Lys-Ala-MCA of 688 m and 11.02 m(-1) s(-1), respectively, and was localized in the periplasm. DPP5 elaborately complemented DPP7 in liberation of dipeptides with hydrophobic P1 residues. Examinations of DPP- and gingipain gene-disrupted mutants indicated that DPP4, DPP5, DPP7, and DPP11 together with Arg- and Lys-gingipains cooperatively liberate most dipeptides from nutrient oligopeptides. This is the first study to report that DPP5 is expressed not only in eukaryotes, but also widely distributed in bacteria and archaea.
• Development of the first internally-quenched fluorescent substrates of human cathepsin C: The application in the enzyme detection in biological samples
  作者：Monika Łęgowska、Yveline Hamon、Anna Wojtysiak、Renata Grzywa、Marcin Sieńczyk、Timo Burster、Brice Korkmaz、Adam Lesner

  DOI：10.1016/j.abb.2016.10.007

  日期：2016.12

  of this study, the first fluorescence resonance energy transfer peptides for the measurement of the activity of human cathepsin C were designed and synthesized. Two series of tetra- and pentapeptide substrates enabled the detailed S' specificity study of cathepsin C, which has been examined for the first time. The extensive enzymatic studies of the obtained compounds resulted in the selection of the

  组织蛋白酶C是一种广泛表达的半胱氨酸外肽酶，主要用于嗜中性粒细胞，细胞毒性T淋巴细胞和肥大细胞中与颗粒相关的促炎性丝氨酸蛋白酶的活化。已经显示出该酶可以在细胞外分泌。然而，其在人体液/生理样品中的发生还没有被彻底研究。在本研究过程中，设计并合成了用于测量人组织蛋白酶C活性的首个荧光共振能量转移肽。两个系列的四肽和五肽底物可以对组织蛋白酶C进行详细的S特异性研究，这是首次进行了研究。对获得的化合物进行的广泛酶研究导致选择了高度特异性和选择性的底物Thi-Ala（Mca）-Ser-Gly-Tyr（3-NO2）-NH2，该酶已成功用于组织蛋白酶C活性的检测在复杂的生物样品中，例如细胞裂解液，尿液和支气管肺泡灌洗液。进行所选底物的分子对接是为了更好地了解组织蛋白酶C活性位点中底物的结合模式。