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细菌光感受器YtvA光循环中的暗恢复速率受细胞环境和水合作用的影响。

The dark recovery rate in the photocycle of the bacterial photoreceptor YtvA is affected by the cellular environment and by hydration.

作者信息

Pennacchietti Francesca, Abbruzzetti Stefania, Losi Aba, Mandalari Carmen, Bedotti Roberta, Viappiani Cristiano, Zanacchi Francesca Cella, Diaspro Alberto, Gärtner Wolfgang

机构信息

Fondazione Istituto Italiano di Tecnologia, Genova, Italy.

Dipartimento di Fisica e Scienze della Terra, Università di Parma, Parma, Italy; NEST, Istituto Nanoscienze, Consiglio Nazionale delle Ricerche, Pisa, Italy.

出版信息

PLoS One. 2014 Sep 11;9(9):e107489. doi: 10.1371/journal.pone.0107489. eCollection 2014.

Abstract

We report thermal recovery kinetics of the lit state into the parental dark state, measured for the blue light-sensing photoreceptor YtvA inside overexpressing E. coli and B. subtilis bacterial cells, performed for the wild type and several mutated proteins. Recovery was followed as a recovery of the fluorescence, as this property is only found for the parental but not for the photochemically generated lit state. When cells were deposited onto a microscope glass plate, the observed thermal recovery rate in the photocycle was found ca. ten times faster in comparison to purified YtvA in solution. When the E. coli or B. subtilis colonies were soaked in an isotonic buffer, the dark relaxation became again much slower and was very similar to that observed for YtvA in solution. The observed effects show that rate constants can be tuned by the cellular environment through factors such as hydration.

摘要

我们报告了亮态向亲本暗态的热恢复动力学,该动力学是针对在过表达的大肠杆菌和枯草芽孢杆菌细胞内的蓝光感应光感受器YtvA进行测量的,测量对象为野生型和几种突变蛋白。恢复过程通过荧光恢复来跟踪,因为这种特性仅在亲本状态下存在,而在光化学产生的亮态中不存在。当细胞沉积在显微镜载玻片上时,发现光循环中的热恢复速率约为溶液中纯化的YtvA的十倍。当大肠杆菌或枯草芽孢杆菌菌落浸泡在等渗缓冲液中时,暗态弛豫再次变得慢得多,并且与溶液中YtvA的弛豫非常相似。观察到的效应表明,速率常数可以通过细胞环境中的因素(如水合作用)进行调节。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b0c4/4161478/dba700b558f5/pone.0107489.g001.jpg

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