(S)-15-azido-2,2-dimethyl-4,12-dioxo-3,13-dioxa-5,11-diazapentadecane-6-carboxylic acis | 1167421-24-0

• 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: (S)-15-azido-2,2-dimethyl-4,12-dioxo-3,13-dioxa-5,11-diazapentadecane-6-carboxylic acis
• 英文别名: (S)-15-azido-2,2-dimethyl-4,12-dioxo-3,13-dioxa-5,11-diazapentadecane-6-carboxylic acid；(2S)-6-(2-azidoethoxycarbonylamino)-2-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid
• CAS: 1167421-24-0
• 化学式: C₁₄H₂₅N₅O₆
• 分子量: 359.382
• InChiKey: YXVLPJSNUYXKCN-JTQLQIEISA-N

计算性质

• 辛醇/水分配系数(LogP)：
  2.3
• 重原子数：
  25
• 可旋转键数：
  13
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.79
• 拓扑面积：
  128
• 氢给体数：
  3
• 氢受体数：
  8

反应信息

• 作为反应物：
  描述：

  (S)-15-azido-2,2-dimethyl-4,12-dioxo-3,13-dioxa-5,11-diazapentadecane-6-carboxylic acis 在 盐酸 、 三氟乙酸 作用下， 以 二氯甲烷 、 水 为溶剂， 反应 0.5h， 生成 N-Ε-炔丙氧基羰基-L-赖氨酸盐酸盐
  参考文献：
  名称：

  Click Strategies for Single-Molecule Protein Fluorescence

  摘要：

  Single-molecule methods have matured into central tools for studies in biology. Foerster resonance energy transfer (FRET) techniques, in particular, have been widely applied to study biomolecular structure and dynamics. The major bottleneck for a facile and general application of these studies arises from the need to label biological samples site-specifically with suitable fluorescent dyes. In this work, we present an optimized strategy combining click chemistry and the genetic encoding of unnatural amino acids (UAAs) to overcome this limitation for proteins. We performed a systematic study with a variety of clickable UAAs and explored their potential for high-resolution single-molecule FRET (smFRET). We determined all parameters that are essential for successful single-molecule studies, such as accessibility of the probes, expression yield of proteins, and quantitative labeling. Our multiparameter fluorescence analysis allowed us to gain new insights into the effects and photophysical properties of fluorescent dyes linked to various UAAs for smFRET measurements. This led us to determine that, from the extended tool set that we now present, genetically encoding propargyllysine has major advantages for state-of-the-art measurements compared to other UAAs. Using this optimized system, we present a biocompatible one-step dual-labeling strategy of the regulatory protein RanBP3 with full labeling position freedom. Our technique allowed us then to determine that the region encompassing two FxFG repeat sequences adopts a disordered but collapsed state. RanBP3 serves here as a prototypical protein that, due to its multiple cysteines, size, and partially disordered structure, is not readily accessible to any of the typical structure determination techniques such as smFRET, NMR, and X-ray crystallography.

  DOI：

  10.1021/ja210587q
• 作为产物：
  描述：

  N-Boc-N^ε-((2-bromoethoxy)carbonyl)-lysine 在 sodium azide 作用下， 以 N,N-二甲基甲酰胺 为溶剂， 反应 48.0h， 生成 (S)-15-azido-2,2-dimethyl-4,12-dioxo-3,13-dioxa-5,11-diazapentadecane-6-carboxylic acis
  参考文献：
  名称：

  Click Strategies for Single-Molecule Protein Fluorescence

  摘要：

  Single-molecule methods have matured into central tools for studies in biology. Foerster resonance energy transfer (FRET) techniques, in particular, have been widely applied to study biomolecular structure and dynamics. The major bottleneck for a facile and general application of these studies arises from the need to label biological samples site-specifically with suitable fluorescent dyes. In this work, we present an optimized strategy combining click chemistry and the genetic encoding of unnatural amino acids (UAAs) to overcome this limitation for proteins. We performed a systematic study with a variety of clickable UAAs and explored their potential for high-resolution single-molecule FRET (smFRET). We determined all parameters that are essential for successful single-molecule studies, such as accessibility of the probes, expression yield of proteins, and quantitative labeling. Our multiparameter fluorescence analysis allowed us to gain new insights into the effects and photophysical properties of fluorescent dyes linked to various UAAs for smFRET measurements. This led us to determine that, from the extended tool set that we now present, genetically encoding propargyllysine has major advantages for state-of-the-art measurements compared to other UAAs. Using this optimized system, we present a biocompatible one-step dual-labeling strategy of the regulatory protein RanBP3 with full labeling position freedom. Our technique allowed us then to determine that the region encompassing two FxFG repeat sequences adopts a disordered but collapsed state. RanBP3 serves here as a prototypical protein that, due to its multiple cysteines, size, and partially disordered structure, is not readily accessible to any of the typical structure determination techniques such as smFRET, NMR, and X-ray crystallography.

  DOI：

  10.1021/ja210587q

文献信息

• MUTANT VIRUS, PREPARATION METHOD THEREFOR AND APPLICATION THEREOF
  申请人：PEKING UNIVERSITY

  公开号：US20200268870A1

  公开（公告）日：2020-08-27

  The present invention relates to a mutated virus. Said virus can be an influenza virus of human or other animal origin. The present invention also relates to a method for preparing the mutated virus, the method comprising introducing UAG codons into positions upstream of the stop codons per se of one or more genes of a viral genome by reverse genetic techniques. The present invention further relates to uses of the mutated virus, for example, as a live attenuated vaccine, or in replication of controllable and safe virus models, and the like.

  本发明涉及一种突变病毒。所述病毒可以是人类或其他动物来源的流感病毒。本发明还涉及一种制备突变病毒的方法，该方法包括通过反向遗传技术将UAG密码子引入一个或多个病毒基因组基因的终止密码子本身的上游位置。本发明进一步涉及突变病毒的使用，例如，作为活疫苗，或用于复制可控制和安全的病毒模型等。
• Methods
  申请人：Nguyen Duy P.

  公开号：US20120077948A1

  公开（公告）日：2012-03-29

  The invention relates to a method of making a polypeptide comprising an orthogonal functional group, said orthogonal functional group being comprised by an aliphatic amino acid or amino acid derivative, said method comprising providing a host cell; providing a nucleic acid encoding the polypeptide of interest; providing a tRNA-tRNA synthetase pair orthogonal to said host cell; adding an amino acid or amino acid derivative comprising the orthogonal functional group of interest, wherein said amino acid or amino acid derivative is a substrate for said orthogonal tRNA synthetase, wherein said amino acid or amino acid derivative has an aliphatic carbon backbone; and incubating to allow incorporation of said amino acid or amino acid derivative into the polypeptide of interest via the orthogonal tRNA-tRNA synthetase pair. The invention also relates to certain amino acids, and to polypeptides comprising same.

  本发明涉及一种制备具有正交功能基团的多肽的方法，所述正交功能基团由脂肪族氨基酸或氨基酸衍生物组成，该方法包括提供宿主细胞；提供编码所需多肽的核酸；提供与所述宿主细胞正交的tRNA-tRNA合成酶对；添加包含所需正交功能基团的氨基酸或氨基酸衍生物，其中所述氨基酸或氨基酸衍生物是正交tRNA合成酶的底物，其中所述氨基酸或氨基酸衍生物具有脂肪族碳骨架；孵育以允许通过正交tRNA-tRNA合成酶对将所述氨基酸或氨基酸衍生物并入所需多肽中。本发明还涉及某些氨基酸以及包含它们的多肽。
• Genetic Encoding and Labeling of Aliphatic Azides and Alkynes in Recombinant Proteins <i>via</i> a Pyrrolysyl-tRNA Synthetase/tRNA_CUA Pair and Click Chemistry
  作者：Duy P. Nguyen、Hrvoje Lusic、Heinz Neumann、Prashant B. Kapadnis、Alexander Deiters、Jason W. Chin

  DOI：10.1021/ja900553w

  日期：2009.7.1

  recombinant proteins in Escherichia coli. Proteins containing the alkyne functional group are labeled with an azido biotin and an azido fluorophore, via copper catalyzed [3+2] cycloaddition reactions, to produce the corresponding triazoles in good yield. The methods reported are useful for the site-specific labeling of recombinant proteins and may be combined with mutually orthogonal methods of introducing

  我们证明了正交 Methanosarcina barkeri MS pyrrolysyl-tRNA 合成酶/tRNA(CUA) 对指导 N6-[(2-丙炔氧基)羰基]-L-赖氨酸的高效、位点特异性掺入，含有碳-碳三键，和 N6-[(2-叠氮乙氧基)羰基]-L-赖氨酸，含有一个叠氮基，在大肠杆菌中转化为重组蛋白。含有炔官能团的蛋白质用叠氮基生物素和叠氮基荧光团标记，通过铜催化的 [3+2] 环加成反应，以良好的产率产生相应的三唑。报道的方法可用于重组蛋白质的位点特异性标记，并且可以与将非天然氨基酸引入蛋白质的相互正交方法以及蛋白质标记的化学正交方法相结合。
• Targeting interleukin-4 to the arthritic joint
  作者：Valerie Spieler、Marie-Gabrielle Ludwig、Janet Dawson、Bruno Tigani、Amanda Littlewood-Evans、Caterina Safina、Hilmar Ebersbach、Klaus Seuwen、Martina Raschig、Björn ter Mors、Thomas D. Müller、Lorenz Meinel、Tessa Lühmann

  DOI：10.1016/j.jconrel.2020.07.005

  日期：2020.10

  Anti-inflammatory cytokines are a promising class of therapeutics for treatment of rheumatoid arthritis (RA), but their use is currently limited by a rapid clearance and systemic toxicity. Interleukin-4 is a small cytokine with potential for RA therapy. To increase its pharmacokinetic features, we engineered a murine IL4 conjugate by incorporating an unnatural amino acid through genetic code expansion to which PEG-folate, as a targeting moiety and PEG alone as control, were site-specifically bound. Both IL4 conjugates retained bioactivity and induced primary murine macrophage polarization into an alternatively activated (M2) related phenotype. The PEGylated conjugates had a terminal half-life of about four hours in healthy mice compared to unPEGylated IL4 (0.76 h). We showed that both conjugates successfully accumulated into arthritic joints in an antigen-induced arthritis (AIA) mouse model, as assessed by non-invasive fluorescence imaging. The modular nature of the IL4 conjugate chemistry presented herein facilitates easy adaption of PEG chain length and targeting moieties for further improvement of half-life and targeting function for future efficacy studies.