携异戊烯基光亲和化学探针的合成和应用

应用“一锅法”耦连反应为关键步骤合成了3个同时含有异戊烯链和叠氮基团的光亲和探针分子。在光照条件下这些化合物和酿酒酵母总蛋白反应后,经过点击反应与含有生物素的报告分子连接,再进行亲和素印迹分析,初步表明它们可作为钓取与异戊烯链相互作用蛋白的化学探针。     

光亲和标记磷脂酸类似物的合成

合成了光敏基团位于sn-1脂肪酰基上的光亲和标记磷脂酸(PA)类似物,选用了有较高C-H插入效率的全氟苯基叠氮化合物作为光敏基团.用酶化学方法在PA类似物中引入了同位素标记³³P.初步实验表明,合成的PA类似物与天然PA一样对cAMP-磷酸二酯酶有激活作用,提示合成的PA类似物可进一步用于该酶的光亲和标记.     

一种侧链液晶高分子的合成与表征

合成了一种主链为甲基丙烯酸酯 ,侧链是三个苯环以酯键相连的介晶基元 ,柔性间隔段为两个亚甲基的新的侧链液晶高分子 .聚合物和单体的化学结构、液晶相转变和液晶态织构采用红外光谱、核磁共振、偏光显微镜、差热分析和广角X光衍射进行了表征 .研究表明 ,单体呈现近晶相和向列相两个液晶相 ,聚合物在很宽的温度范围内呈向列相 .该单体和聚合物在一定条件下可形成固化诱导条带织构     

GABA门控的氯离子通道光亲和探针的设计与合成

为了研究GABA门控的氯离子通道上杀虫活性化合物的结合位点， 以5e-叔丁基-2e-(4-取代丙炔基苯基)-1,3-二噻烷为先导，设计和合成了作用于GABA门控的氯离子通道上非竞争结合位点的新光亲和探针化合物。通过³H EBOB法测试， 新探针化合物的最大半抑制浓度小于35nM， 能够在300nm的紫外光的照射下迅速发生光反应。如果在这些化合物中引入同位素碘，可望成为好的光亲和标记探针。     

新型侧链液晶聚氨酯扩链剂的合成与表征

本文合成了用以制备侧链型液晶聚氨酯的扩链剂二醇－４（４－甲氧基苯酰氧基）苯甲酸－双（β－羟乙基）氨基乙酯（ＩＶ），并以ＨＮＭＲ。ＦＴＩＲ、元素分析等测试手段对此二醇进行了表征，测试结果与设定结构相一致     

一种近红外荧光探针的合成、表征及应用

合成了一种卟啉类小分子荧光探针5,10,15,20-四对羟苯基卟啉(THPP)并对其结构进行表征.通过荧光光谱法,可以分别定性、定量检测铜离子(Cu2+)、焦磷酸根(PPi)及碱性磷酸酶(ALP).研究结果可以转化为一个代表性综合实验,激发学生对科学研究的极大兴趣,培养学生利用综合知识的能力.实验囊括了有机化学、分析化学、生物化学及无机化学等多个学科的多个知识点及多种重要的仪器操作技术,有助于学生综合实验能力的提升.     

聚丙烯酸酯侧链液晶聚合物的合成与表征

以对羟基苯甲酸、氯乙醇和丙烯酸为主要原料,经醚化、酯化和酰氯化反应合成了中间体和含液晶基元的丙烯酸酯单体,后者经自由基聚合合成了聚丙烯酸酯侧链液晶聚合物。用偏光显微镜观察了单体和聚合物的织态结构,用DSC和IR对聚合物进行了表征。结果表明,单体和聚合物均呈现向列型液晶织态结构,聚合物在较宽的温度范围内有很好的液晶性。     

一种西佛碱侧链型液晶高分子的合成及表征

一种西佛碱侧链型液晶高分子的合成及表征邹友思，林国良，姚青青，戴李宗，潘容华（厦门大学化学系厦门３６１００５）关键词联苯，西佛碱，侧链型，液晶高分子，基团转移聚合Ｆｉｎｋｅｌｌｎａｎｎ等提出的柔性基去偶合概念’］已在侧链型液晶高分子的研究中得到了证实...     

一种侧链含偶氮基团共聚物的合成及其多层膜的制备与表征

环氧氯丙烷与4,4′-二羟基偶氮苯(AZO)经缩聚反应合成了预聚物DGAZO(聚4,4′-二缩水甘油醚偶氮苯); DGAZO通过开环加成反应与PEI(枝化聚乙烯亚胺)连接制得侧链含偶氮基团的共聚物DGAZO-co-PEI(聚4,4′-二缩水甘油醚偶氮苯-co-聚乙烯亚胺),其结构经UV,IR和荧光光谱表征.DGAZO-...     

一种新的荧光素类荧光探针的设计合成及其对血清中组氨酸的选择性标记

组氨酸是人类必需的一种氨基酸 .它含有高反应活性的咪唑环 ,并在生物体系中金属离子的传输和神经递质的传送等方面起着重要的作用 [1,2 ] .所以 ,生物体液中组氨酸的选择性测定对生物化学的研究具有重要意义 .然而 ,由于各种氨基酸在结构及性质上的相似性 ,选择性测定单独的氨基酸常常需要借助如高效液相色谱和毛细管电泳等一些分离技术才能完成 .各种化合物的官能团反应可为一些高选择性的分子识别提供基础[3] .目前各种主体分子 ,包括荧光纳米探针和分子印迹聚合物等在生物分子的选择性识别方面发挥着重要的作用[4～ 6 ] .环氧氯丙烷作为…     

Design and Synthesis of Photoaffinity Probe Candidates for the GABA-gated Chloride Channel

In order to characterize binding sites of insecticidal compounds on GABA gated chloride channel,new photoaf-finity probe candidates based on 5e-t-butyl-2e-[4-(substituted-propynyl)phenyl]-1,3-dithiane for the noncompetitiveblocker(NCB)site of the γ-aminobutyric acid(GABA)-gated chloride channel were designed and synthesized,andtheir potency as an inhibitor on NCB was measured by 4'-ethynyl-4-n-[2,3-~3H_2]-propylbicycloorthobenzoate(~3HEBOB)assay.The synthesized compounds showed high inhibition activities with half maximum inhibition concen-trations(IC_(50))of lower than 35 nmol/L and were very stable in binding conditions as well photoreacted quickly at300 nm light.These new compounds are expected to be good photoaffinity labeling probes if radioisotope iodine isincorporated.     

Cross-linking chemistry and biology: development of multifunctional photoaffinity probes

An efficient method of photoaffinity labeling has been developed based on rationally designed multifunctional photoprobes. Photoaffinity techniques have been used to elucidate the protein structure at the interface of biomolecules by the photochemical labeling of interacting sites. However, the identification of labeled sites within target proteins is often difficult. Novel biotinyl bioprobes bearing a diazirine photophore have contributed significantly to the rapid elucidation of ligand binding sites within proteins, thereby extending conventional photoaffinity methods. This article discusses the synthesis and applications of various photoprobes bearing a biotin, including strategies using cleavable linkages between photophores. The combination of photoaffinity methods with chip technology is also described as a novel entry to rapid affinity-based screening of inhibitors. This review focuses on a rapid and reliable photoaffinity method utilizing diazirine-based multifunctional photoprobes with numerous potential applications in functional proteomics of biomolecular interactions.     

Recent Applications of Diazirines in Chemical Proteomics

The elucidation of substrate–protein interactions is an important component of the drug development process. Due to the complexity of native cellular environments, elucidating these fundamental biochemical interactions remains challenging. Photoaffinity labeling (PAL) is a versatile technique that can provide insight into ligand-target interactions. By judicious modification of substrates with a photoreactive group, PAL creates a covalent crosslink between a substrate and its biological target following UV-irradiation. Among the commonly employed photoreactive groups, diazirines have emerged as the gold standard. In this Minireview, recent developments in the field of diazirine-based photoaffinity labeling will be discussed, with emphasis being placed on their applications in chemical proteomic studies.     

Unravelling the Allosteric Targeting of PHGDH at the ACT-Binding Domain with a Photoactivatable Diazirine Probe and Mass Spectrometry Experiments

The serine biosynthetic pathway is a key element contributing to tumor proliferation. In recent years, targeting of phosphoglycerate dehydrogenase (PHGDH), the first enzyme of this pathway, intensified and revealed to be a promising strategy to develop new anticancer drugs. Among attractive PHGDH inhibitors are the α-ketothioamides. In previous work, we have demonstrated their efficacy in the inhibition of PHGDH in vitro and in cellulo. However, the precise site of action of this series, which would help the rational design of new inhibitors, remained undefined. In the present study, the detailed mechanism-of-action of a representative α-ketothioamide inhibitor is reported using several complementary experimental techniques. Strikingly, our work led to the identification of an allosteric site on PHGDH that can be targeted for drug development. Using mass spectrometry experiments and an original α-ketothioamide diazirine-based photoaffinity probe, we identified the 523Q-533F sequence on the ACT regulatory domain of PHGDH as the binding site of α-ketothioamides. Mutagenesis experiments further documented the specificity of our compound at this allosteric site. Our results thus pave the way for the development of new anticancer drugs using a completely novel mechanism-of-action.     

Design and Synthesis of Matrix Metalloprotease Photoaffinity Trimodular Probes

To explore the molecular mechanism of the matrix metalloproteinases (MMPs) in tumor processes, two photoaffinity trimodular probes were designed and synthesized based on the structure activity relationship and the following photoaffinity labelling experiments afforded positive results.     

Exploiting Benzophenone Photoreactivity To Probe the Phospholipid Environment and Insertion Depth of the Cell-Penetrating Peptide Penetratin in Model Membranes

Penetratin (RQIKIWFQNRRMKWKK) enters cells by different mechanisms, including membrane translocation, thus implying that the peptide interacts with the lipid bilayer. Penetratin also crosses the membrane of artificial vesicles, depending on their phospholipid content. To evaluate the phospholipid preference of penetratin, as the first step of translocation, we exploited the benzophenone triplet kinetics of hydrogen abstraction, which is slower for secondary than for allylic hydrogen atoms. By using multilamellar vesicles of varying phospholipid content, we identified and characterized the cross-linked products by MALDI-TOF mass spectrometry. Penetratin showed a preference for negatively charged (vs. zwitterionic) polar heads, and for unsaturated (vs. saturated) and short (vs. long) saturated phospholipids. Our study highlights the potential of using benzophenone to probe the environment and insertion depth of membranotropic peptides in membranes.