000111840 001__ 111840 000111840 005__ 20180211172522.0 000111840 0247_ $$2pmid$$apmid:22834887 000111840 0247_ $$2DOI$$a10.1021/jp2103512 000111840 0247_ $$2WOS$$aWOS:000307749100001 000111840 037__ $$aPreJuSER-111840 000111840 041__ $$aeng 000111840 082__ $$a530 000111840 084__ $$2WoS$$aChemistry, Physical 000111840 1001_ $$0P:(DE-Juel1)VDB109632$$aSeemann, Klaus M.$$b0$$uFZJ 000111840 245__ $$aOptical pH Detection within a Protein Crystal 000111840 260__ $$aWashington, DC$$bSoc.$$c2012 000111840 300__ $$a9873-9881 000111840 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article 000111840 3367_ $$2DataCite$$aOutput Types/Journal article 000111840 3367_ $$00$$2EndNote$$aJournal Article 000111840 3367_ $$2BibTeX$$aARTICLE 000111840 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000111840 3367_ $$2DRIVER$$aarticle 000111840 440_0 $$03694$$aJournal of Physical Chemistry B$$v116$$x1520-6106$$y33 000111840 500__ $$3POF3_Assignment on 2016-02-29 000111840 500__ $$aRecord converted from VDB: 16.11.2012 000111840 520__ $$aThe pH is one of the key parameters governing protein conformation and activity. In protein crystals, however, the pH is so far not accessible by experiment. Here, we report on the optical detection of the pH in a lysozyme crystal employing the pH-sensitive fluorescent dyes SNARF-1 and SNARF-4F. The molecular probes were loaded into the crystal by diffusion. Two-dimensional fluorescence spectra of the labeled protein crystal were recorded, and the average pH of the crystal at different bath pH's was determined by calibrating fluorescence peak ratios. In addition, we used two-photon microscopy to spatially resolve the pH inside a lysozyme crystal three-dimensionally and to follow pH changes in response to a pH change of the bath over time. At equilibrium at bath pH between 5.5 and 8.0, we found a pH in the water-filled crystal channels that was ΔpH = -0.3 to -1.0 lower than that of the bath. This corresponds to a 2- to 10-fold higher proton concentration in the crystal channels than in the bath. The lower pH at equilibrium in the crystal channels can be explained by slower proton diffusion in the channels than in the bath and a resulting proton accumulation in the crystal for conservation of mass and so an equilibrium of proton flux. 000111840 536__ $$0G:(DE-Juel1)FUEK412$$2G:(DE-HGF)$$aGrundlagen für zukünftige Informationstechnologien$$cP42$$x0 000111840 588__ $$aDataset connected to Web of Science, Pubmed 000111840 650_7 $$2WoSType$$aJ 000111840 7001_ $$0P:(DE-HGF)0$$aKiefersauer, Reiner$$b1 000111840 7001_ $$0P:(DE-HGF)0$$aJacob, Uwe$$b2 000111840 7001_ $$0P:(DE-HGF)0$$aKuhn, Bernd$$b3 000111840 773__ $$0PERI:(DE-600)2006039-7$$a10.1021/jp2103512$$gVol. 116, p. 9873-9881$$p9873-9881$$q116<9873-9881$$tThe @journal of physical chemistry <Washington, DC> / B$$v116$$x1520-6106$$y2012 000111840 8567_ $$uhttp://dx.doi.org/10.1021/jp2103512 000111840 909CO $$ooai:juser.fz-juelich.de:111840$$pVDB 000111840 9131_ $$0G:(DE-Juel1)FUEK412$$1G:(DE-HGF)POF2-420$$2G:(DE-HGF)POF2-400$$bSchlüsseltechnologien$$kP42$$lGrundlagen für zukünftige Informationstechnologien (FIT)$$vGrundlagen für zukünftige Informationstechnologien$$x0 000111840 9132_ $$0G:(DE-HGF)POF3-529H$$1G:(DE-HGF)POF3-520$$2G:(DE-HGF)POF3-500$$aDE-HGF$$bKey Technologies$$lFuture Information Technology - Fundamentals, Novel Concepts and Energy Efficiency (FIT)$$vAddenda$$x0 000111840 9141_ $$y2012 000111840 915__ $$0StatID:(DE-HGF)0010$$2StatID$$aJCR/ISI refereed 000111840 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR 000111840 915__ $$0StatID:(DE-HGF)0110$$2StatID$$aWoS$$bScience Citation Index 000111840 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded 000111840 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection 000111840 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bThomson Reuters Master Journal List 000111840 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS 000111840 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline 000111840 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database 000111840 915__ $$0StatID:(DE-HGF)1020$$2StatID$$aDBCoverage$$bCurrent Contents - Social and Behavioral Sciences 000111840 9201_ $$0I:(DE-Juel1)PGI-6-20110106$$gPGI$$kPGI-6$$lElektronische Eigenschaften$$x0 000111840 970__ $$aVDB:(DE-Juel1)140351 000111840 980__ $$aVDB 000111840 980__ $$aConvertedRecord 000111840 980__ $$ajournal 000111840 980__ $$aI:(DE-Juel1)PGI-6-20110106 000111840 980__ $$aUNRESTRICTED