OREGON GREEN 488-Carboxylic acid-maleimide

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 苯并吡喃
• 英文名称: OREGON GREEN 488-Carboxylic acid-maleimide
• 英文别名: OG NHS ester；Oregon Green 488 carboxylic acid, succinimidyl ester, 6-isomer；Difluorocarboxyfluorescein NHS Ester, 6-isomer；2-(2,7-difluoro-3-hydroxy-6-oxoxanthen-9-yl)-4-(2,5-dioxopyrrolidin-1-yl)oxycarbonylbenzoic acid
• 化学式: C₂₅H₁₃F₂NO₉
• 分子量: 509.376
• InChiKey: AXNGAJYYONSYCY-UHFFFAOYSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  1.8
• 重原子数：
  37
• 可旋转键数：
  5
• 环数：
  5.0
• sp3杂化的碳原子比例：
  0.08
• 拓扑面积：
  148
• 氢给体数：
  2
• 氢受体数：
  11

反应信息

• 作为反应物：
  描述：

  OREGON GREEN 488-Carboxylic acid-maleimide 、 N,N-双(羧甲基)-L-赖氨酸 在 N,N-二异丙基乙胺 作用下， 以 N,N-二甲基甲酰胺 为溶剂， 反应 2.0h， 生成
  参考文献：
  名称：

  Specific and Stable Fluorescence Labeling of Histidine-Tagged Proteins for Dissecting Multi-Protein Complex Formation

  摘要：

  Labeling of proteins with fluorescent dyes offers powerful means for monitoring protein interactions in vitro and in live cells. Only a few techniques for noncovalent fluorescence labeling with well-defined localization of the attached dye are currently available. Here, we present an efficient method for site-specific and stable noncovalent fluorescence labeling of histidine-tagged proteins. Different fluorophores were conjugated to a chemical recognition unit bearing three NTA moieties (tris-NTA). In contrast to the transient binding of conventional mono-NTA, the multivalent interaction of tris-NTA conjugated fluorophores with oligohistidine-tagged proteins resulted in complex lifetimes of more than an hour. The high selectivity of tris-NTA toward cumulated histidines enabled selective labeling of proteins in cell lysates and on the surface of live cells. Fluorescence labeling by tris-NTA conjugates was applied for the analysis of a ternary protein complex in solution and on surfaces. Formation of the complex and its stoichiometry was studied by analytical size exclusion chromatography and fluorescence quenching. The individual interactions were dissected on solid supports by using simultaneous mass-sensitive and multicolor fluorescence detection. Using these techniques, formation of a 1:1:1 stoichiometry by independent interactions of the receptor subunits with the ligand was shown. The incorporation of transition metal ions into the labeled proteins upon labeling with tris-NTA fluorophore conjugates provided an additional sensitive spectroscopic reporter for detecting and monitoring protein-protein interactions in real time. A broad application of these fluorescence conjugates for protein interaction analysis can be envisaged.

  DOI：

  10.1021/ja0563105

文献信息

• Condensation of 2-((Alkylthio)(aryl)methylene)malononitrile with 1,2-Aminothiol as a Novel Bioorthogonal Reaction for Site-Specific Protein Modification and Peptide Cyclization
  作者：Xiaoli Zheng、Zhuoru Li、Wei Gao、Xiaoting Meng、Xuefei Li、Louis Y. P. Luk、Yibing Zhao、Yu-Hsuan Tsai、Chuanliu Wu

  DOI：10.1021/jacs.9b11875

  日期：2020.3.18

  2-aminothiol functionality can be introduced into a peptide or a protein as an N-terminal cysteine or an unnatural amino acid. The bioorthogonality of this reaction was demonstrated by site-specific labeling of not only synthetic peptides and a purified recombinant protein but also proteins on mammalian cells and phages. Unlike other reagents in bioorthogonal reactions, the chemical and physical properties of

  肽和蛋白质的位点特异性修饰在探测和扰动生物系统中具有广泛的应用。在此我们报告了 1,2-氨基硫醇可以在生物相容性条件下与 2-((烷硫基)(芳基)亚甲基)丙二腈 (TAMM) 快速、特异性和有效地反应。该反应经历了一种独特的机制，包括硫醇-乙烯基硫化物交换、环化和二氰基甲烷的消除，以形成 2-芳基-4,5-二氢噻唑 (ADT) 作为稳定的产物。可以将 1,2-氨基硫醇官能团作为 N 端半胱氨酸或非天然氨基酸引入肽或蛋白质中。该反应的生物正交性通过不仅合成肽和纯化的重组蛋白而且哺乳动物细胞和噬菌体上的蛋白质的位点特异性标记来证明。与生物正交反应中的其他试剂不同，TAMM 的化学和物理特性可以轻松调整。TAMM 还可用于生成基于噬菌体的环肽文库，而不会降低噬菌体的感染性。使用这种方法，我们鉴定了对不同蛋白质靶标具有高亲和力的 ADT 环肽，为生物学研究和潜在治疗提供了有价值的工具。此外，TAMM
• IMAGING AND THERAPEUTIC METHOD USING PROGENITOR CELLS
  申请人：Low Philip Stewart

  公开号：US20100226967A1

  公开（公告）日：2010-09-09

  The invention relates to a method of treating or diagnosing a disease state worsened by progenitor cells. The method utilizes a composition comprising a conjugate or complex of the general formula A b -X wherein the group A b comprises a vitamin, or an analog thereof, that binds to progenitor cells, and when the conjugate is being used for treatment of the disease state, the group X comprises an antigen, a cytotoxin, or a compound capable of altering progenitor cell function, and when the conjugate is being used for diagnosing the disease state, the group X comprises a quantifiable marker.

  本发明涉及一种治疗或诊断由祖细胞恶化引起的疾病状态的方法。该方法利用包含通式Ab-X的共轭物或复合物的组合物，其中Ab组包括维生素或其类似物，该维生素或其类似物与祖细胞结合。当该共轭物用于治疗疾病状态时，组X包括抗原、细胞毒素或能够改变祖细胞功能的化合物；当该共轭物用于诊断疾病状态时，组X包括可量化的标记物。
• METHOD OF DETECTING ENDOTHELIAL PROGENITOR CELLS
  申请人：PURDUE RESEARCH FOUNDATION

  公开号：US20130266964A1

  公开（公告）日：2013-10-10

  A method of detecting endothelial progenitor cells is provided. The method involves contacting a population of cells with a composition comprising a conjugate or complex of the formula A b -X where the group A b comprises a folate that binds to endothelial progenitor cells, and the group X comprises a fluorescent chromophore, and eliciting a fluorescent response from bound A b -X.

  提供了一种检测内皮祖细胞的方法。该方法涉及将细胞群体与包含式Ab-X的共轭物或复合物的组合物接触，其中组Ab包括结合内皮祖细胞的叶酸，组X包括荧光色团，并从结合的Ab-X中引出荧光响应。
• Synthesis and in vitro evaluation of cyclic NGR peptide targeted thermally sensitive liposome
  作者：Ayele H. Negussie、Jenna L. Miller、Goutham Reddy、Steven K. Drake、Bradford J. Wood、Matthew R. Dreher

  DOI：10.1016/j.jconrel.2009.12.031

  日期：2010.4

  The Asn-Gly-Arg (NGR) motif in both cyclic and linear form has previously been shown to specifically bind to CD13/aminopeptidase N that is selectively overexpressed in tumor vasculature and some tumor cells. However, previous versions of cyclic NGR used a liable disulfide bridge between cysteine residues that may be problematic for liposome targeting due to disulfide bond formation between adjacent peptides on the liposomal surface. In this study, we report the design, synthesis, and characterization of a novel cyclic NGR-containing peptide, cKNGRE, which does not contain a disulfide bridge. cKNGRE was synthesized in good yield and purity and attached to the fluorescent reporter Oregon Green (cKNGRE-OG) and lysolipid-containing temperature sensitive liposomes (LTSLs). The identity of cKNGRE was verified with NMR and mass spectral techniques. In vitro fluorescence microscopy evaluation of cKNGRE-OG demonstrated binding and active uptake by CD13(+) cancer cells and minimal binding to CD13(-) cancer cells. The cKNGRE-OG ligand displayed 3.6-fold greater affinity for CD13(+) cancer cells than a linear NGR-containing peptide. Affinity for CD13(+) cancer cells was similarly improved 10-fold for both the cyclic and linear NGR when presented in a multivalent fashion on the surface of an LTSL cKNGRE-targeted LTSLs rapidly released (>75% in <4 s) doxorubicin at 41.3 degrees C with minimal release at 37 degrees C. These results demonstrate the ability to synthesize a cKNGRE-targeted temperature sensitive liposome that lacks a disulfide bridge and has sufficient binding affinity for biological applications. Published by Elsevier B.V.
• FLUORINATED XANTHENE DERIVATIVES
  申请人：Molecular Probes, Inc.

  公开号：EP0853647B2

  公开（公告）日：2011-06-15