{"title":"荧光染料8-苯胺萘磺酸与大肠杆菌色氨酸合成酶的天然和压力解离- 2二聚体的结合。","authors":"T Seifert, P Bartholmes, R Jaenicke","doi":"10.1515/znc-1984-9-1023","DOIUrl":null,"url":null,"abstract":"<p><p>The beta 2-dimer of tryptophan synthase from Escherichia coli exhibits weak binding of 8-anilinonaphthalene-1-sulfonic acid (ANS). Titrating the dye at 0.2 mM concentration with the apo-beta 2-dimer at atmospheric pressure causes increased fluorescence emission at 480 nm (lambda exc = 380 nm), corresponding to unspecific binding of the ligand to hydrophobic residues. Increasing hydrostatic pressure affects ANS binding. Up to 700 bar, a sigmoidal increase of ANS fluorescence reflects an increase in hydrophobic surface area, probably caused by subunit dissociation. At approximately 1 kbar, a maximum is reached; beyond this value, pressure competes with ligand binding causing fluorescence emission to be decreased again. Pressure release leads to a drastic fluorescence enhancement, ascribed to ANS binding to the partially and reversibly denatured enzyme. Plotting the total fluorescence enhancement vs. pressure yields a profile which parallels the pressure dependent dimer in equilibrium monomer transition monitored by subunit hybridization (T. Seifert, P. Bartholmes, and R. Jaenicke, Biochemistry, in press).</p>","PeriodicalId":23914,"journal":{"name":"Zeitschrift fur Naturforschung. Section C, Biosciences","volume":"39 9-10","pages":"1008-11"},"PeriodicalIF":0.0000,"publicationDate":"1984-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/znc-1984-9-1023","citationCount":"2","resultStr":"{\"title\":\"Binding of the fluorescent dye 8-anilinonaphthalene 1-sulfonic acid to the native and pressure dissociated beta 2-dimer of tryptophan synthase from Escherichia coli.\",\"authors\":\"T Seifert, P Bartholmes, R Jaenicke\",\"doi\":\"10.1515/znc-1984-9-1023\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The beta 2-dimer of tryptophan synthase from Escherichia coli exhibits weak binding of 8-anilinonaphthalene-1-sulfonic acid (ANS). Titrating the dye at 0.2 mM concentration with the apo-beta 2-dimer at atmospheric pressure causes increased fluorescence emission at 480 nm (lambda exc = 380 nm), corresponding to unspecific binding of the ligand to hydrophobic residues. Increasing hydrostatic pressure affects ANS binding. Up to 700 bar, a sigmoidal increase of ANS fluorescence reflects an increase in hydrophobic surface area, probably caused by subunit dissociation. At approximately 1 kbar, a maximum is reached; beyond this value, pressure competes with ligand binding causing fluorescence emission to be decreased again. Pressure release leads to a drastic fluorescence enhancement, ascribed to ANS binding to the partially and reversibly denatured enzyme. Plotting the total fluorescence enhancement vs. pressure yields a profile which parallels the pressure dependent dimer in equilibrium monomer transition monitored by subunit hybridization (T. Seifert, P. Bartholmes, and R. Jaenicke, Biochemistry, in press).</p>\",\"PeriodicalId\":23914,\"journal\":{\"name\":\"Zeitschrift fur Naturforschung. Section C, Biosciences\",\"volume\":\"39 9-10\",\"pages\":\"1008-11\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1984-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1515/znc-1984-9-1023\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Zeitschrift fur Naturforschung. Section C, Biosciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1515/znc-1984-9-1023\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zeitschrift fur Naturforschung. Section C, Biosciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1515/znc-1984-9-1023","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Binding of the fluorescent dye 8-anilinonaphthalene 1-sulfonic acid to the native and pressure dissociated beta 2-dimer of tryptophan synthase from Escherichia coli.
The beta 2-dimer of tryptophan synthase from Escherichia coli exhibits weak binding of 8-anilinonaphthalene-1-sulfonic acid (ANS). Titrating the dye at 0.2 mM concentration with the apo-beta 2-dimer at atmospheric pressure causes increased fluorescence emission at 480 nm (lambda exc = 380 nm), corresponding to unspecific binding of the ligand to hydrophobic residues. Increasing hydrostatic pressure affects ANS binding. Up to 700 bar, a sigmoidal increase of ANS fluorescence reflects an increase in hydrophobic surface area, probably caused by subunit dissociation. At approximately 1 kbar, a maximum is reached; beyond this value, pressure competes with ligand binding causing fluorescence emission to be decreased again. Pressure release leads to a drastic fluorescence enhancement, ascribed to ANS binding to the partially and reversibly denatured enzyme. Plotting the total fluorescence enhancement vs. pressure yields a profile which parallels the pressure dependent dimer in equilibrium monomer transition monitored by subunit hybridization (T. Seifert, P. Bartholmes, and R. Jaenicke, Biochemistry, in press).