P. Borden, Peng Zhang, Amol V. Shivange, J. Marvin, Joseph Cichon, Chuntao Dan, Kaspar Podgorski, Antonio Figueiredo, O. Novák, M. Tanimoto, E. Shigetomi, Mark A. Lobas, Hyun-Tae Kim, Paula K Zhu, Yajun Zhang, W. Zheng, Chengcheng Fan, Guangfu Wang, Bowen Xiang, Lihua Gan, Guang-Xian Zhang, Kaiming Guo, Li Lin, Yuan Cai, Andrew G Yee, Abhi Aggarwal, Huan Bao, Xiaochu Lou, E. Chapman, C. P. Ford, D. Rees, D. Dietrich, B. Khakh, J. Dittman, W. Gan, Minoru Koyama, V. Jayaraman, J. Cheer, H. Lester, J. J. Zhu, L. Looger
Here we design and optimize a genetically encoded fluorescent indicator, iAChSnFR, for the ubiquitous neurotransmitter acetylcholine, based on a bacterial periplasmic binding protein. iAChSnFR shows large fluorescence changes, rapid rise and decay kinetics, and insensitivity to most cholinergic drugs. iAChSnFR revealed large transients in a variety of slice and in vivo preparations in mouse, fish, fly and worm. iAChSnFR will be useful for the study of acetylcholine in all organisms.
{"title":"A Fast Genetically Encoded Fluorescent Sensor for Faithful in vivo Acetylcholine Detection in Mice, Fish, Worms and Flies","authors":"P. Borden, Peng Zhang, Amol V. Shivange, J. Marvin, Joseph Cichon, Chuntao Dan, Kaspar Podgorski, Antonio Figueiredo, O. Novák, M. Tanimoto, E. Shigetomi, Mark A. Lobas, Hyun-Tae Kim, Paula K Zhu, Yajun Zhang, W. Zheng, Chengcheng Fan, Guangfu Wang, Bowen Xiang, Lihua Gan, Guang-Xian Zhang, Kaiming Guo, Li Lin, Yuan Cai, Andrew G Yee, Abhi Aggarwal, Huan Bao, Xiaochu Lou, E. Chapman, C. P. Ford, D. Rees, D. Dietrich, B. Khakh, J. Dittman, W. Gan, Minoru Koyama, V. Jayaraman, J. Cheer, H. Lester, J. J. Zhu, L. Looger","doi":"10.2139/ssrn.3554080","DOIUrl":"https://doi.org/10.2139/ssrn.3554080","url":null,"abstract":"Here we design and optimize a genetically encoded fluorescent indicator, iAChSnFR, for the ubiquitous neurotransmitter acetylcholine, based on a bacterial periplasmic binding protein. iAChSnFR shows large fluorescence changes, rapid rise and decay kinetics, and insensitivity to most cholinergic drugs. iAChSnFR revealed large transients in a variety of slice and in vivo preparations in mouse, fish, fly and worm. iAChSnFR will be useful for the study of acetylcholine in all organisms.","PeriodicalId":286484,"journal":{"name":"BiochemRN: Other Biochemistry Methods (Topic)","volume":"154 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134507026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Glycoproteins play an important role in biological processes such as protein folding, information transmission, nerve conduction, and molecular recognition. Therefore, it is of great significance to design and develop an adsorbent with high adsorption capacity for glycoprotein. In this paper, a novel boronate affinity material (Zr-MOF@S-S@B) was prepared by combining Zr-MOF, disulfide covalent bond (-S-S-), 4-mercaptophenylboronic acid (4-MPBA) for selective enrichment of glycoprotein and elution of glycoprotein at physiological pH. The affinity between boric acid and cis - diol makes the composite material has a high adsorption capacity of 625.5 mg g -1 for ovalbumin (OVA). At the same time, the Zr-MOF@S-S@B has exhibited great reusability and large specific surface area. Furthermore, the material contains disulfide bonds that can release surface binding glycoprotein specifically under physiological pH conditions, effectively avoiding the reduction of glycoprotein activity. In general, the method is successfully used for the enrichment of OVA in egg white samples, which has broad application prospects in the study of glycoprotein.
糖蛋白在蛋白质折叠、信息传递、神经传导和分子识别等生物过程中发挥着重要作用。因此，设计和研制对糖蛋白具有高吸附能力的吸附剂具有重要意义。本文将Zr-MOF、二硫共价键(- s - s -)、4-巯基苯基硼酸(4-MPBA)结合制备了一种新型硼酸亲和材料(Zr-MOF@S-S@B)，可在生理ph下选择性富集糖蛋白并洗脱糖蛋白。硼酸与顺式二醇的亲和关系使该复合材料对卵清蛋白(OVA)具有625.5 mg g -1的高吸附容量。同时，Zr-MOF@S-S@B具有较高的可重用性和较大的比表面积。此外，该材料含有二硫键，可以在生理pH条件下特异性释放表面结合的糖蛋白，有效避免糖蛋白活性的降低。总的来说，该方法成功地用于蛋清样品中OVA的富集，在糖蛋白研究中具有广阔的应用前景。
{"title":"Preparation of Boronate Affinity - Functionalized Metal-Organic Framework Material For Selective Recognition And Separation of Glycoprotein Under Physiological pH","authors":"Baoyue Zhang, Jianghua He, Bailin Guo, Xue Chen, Sheng Bi, Feng Zhang, M. Tian","doi":"10.2139/ssrn.3932111","DOIUrl":"https://doi.org/10.2139/ssrn.3932111","url":null,"abstract":"Glycoproteins play an important role in biological processes such as protein folding, information transmission, nerve conduction, and molecular recognition. Therefore, it is of great significance to design and develop an adsorbent with high adsorption capacity for glycoprotein. In this paper, a novel boronate affinity material (Zr-MOF@S-S@B) was prepared by combining Zr-MOF, disulfide covalent bond (-S-S-), 4-mercaptophenylboronic acid (4-MPBA) for selective enrichment of glycoprotein and elution of glycoprotein at physiological pH. The affinity between boric acid and cis - diol makes the composite material has a high adsorption capacity of 625.5 mg g -1 for ovalbumin (OVA). At the same time, the Zr-MOF@S-S@B has exhibited great reusability and large specific surface area. Furthermore, the material contains disulfide bonds that can release surface binding glycoprotein specifically under physiological pH conditions, effectively avoiding the reduction of glycoprotein activity. In general, the method is successfully used for the enrichment of OVA in egg white samples, which has broad application prospects in the study of glycoprotein.","PeriodicalId":286484,"journal":{"name":"BiochemRN: Other Biochemistry Methods (Topic)","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130022250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. K. Kannuchamy, K. Ramisetty, K. Renukadevi, Rama Krishna Gamidi, C. Heffernan, Andrew A. Stewart, Jian Guo, Gadipelli Srinivas, D. Brett, E. Favvas, Å. Rasmuson
Crystallization experiments performed with highly supercooled solutions produced highly pure (> 99 wt%) and highly crystalline mesocrystals of curcumin from impure solution (~22% of two structurally similar impurities) in one step. These mesocrystals exhibited a crystallographic hierarchy and were composed of perfectly or imperfectly aligned nanometer thick crystallites. X-ray diffraction and spectroscopic analysis confirmed that the spherulites are a new solid form of curcumin. A theoretical hypothesis based on particle aggregation, double nucleation and a repeated secondary nucleation is proposed to explain the spherulite formation mechanism. The experimental results provide for the first time evidence for an organic molecule to naturally form spherulites without the presence of any stabilizing agent. Control experiments performed with highly supercooled pure solutions produced spherulites confirming the formation of spherulites is attributed to the high degree of supercooling and not due to the presence of impurities. Likewise control experiments performed with a lower degree of supercooling produced impure crystals of curcumin via the classical molecular addition mechanisms. These experimental observations all together provide first time evidence for particle mediated crystallization as an alternate and efficient method to purify organic compounds.
{"title":"Pure Curcumin Spherulites from Impure Solution Via Non-Classical Crystallization","authors":"V. K. Kannuchamy, K. Ramisetty, K. Renukadevi, Rama Krishna Gamidi, C. Heffernan, Andrew A. Stewart, Jian Guo, Gadipelli Srinivas, D. Brett, E. Favvas, Å. Rasmuson","doi":"10.2139/ssrn.3844727","DOIUrl":"https://doi.org/10.2139/ssrn.3844727","url":null,"abstract":"Crystallization experiments performed with highly supercooled solutions produced highly pure (> 99 wt%) and highly crystalline mesocrystals of curcumin from impure solution (~22% of two structurally similar impurities) in one step. These mesocrystals exhibited a crystallographic hierarchy and were composed of perfectly or imperfectly aligned nanometer thick crystallites. X-ray diffraction and spectroscopic analysis confirmed that the spherulites are a new solid form of curcumin. A theoretical hypothesis based on particle aggregation, double nucleation and a repeated secondary nucleation is proposed to explain the spherulite formation mechanism. The experimental results provide for the first time evidence for an organic molecule to naturally form spherulites without the presence of any stabilizing agent. Control experiments performed with highly supercooled pure solutions produced spherulites confirming the formation of spherulites is attributed to the high degree of supercooling and not due to the presence of impurities. Likewise control experiments performed with a lower degree of supercooling produced impure crystals of curcumin via the classical molecular addition mechanisms. These experimental observations all together provide first time evidence for particle mediated crystallization as an alternate and efficient method to purify organic compounds.","PeriodicalId":286484,"journal":{"name":"BiochemRN: Other Biochemistry Methods (Topic)","volume":"88 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129621335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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