Department of Neurosciences, Center for Research in Biological Systems, University of California San Diego School of Medicine, La Jolla, California, United States of America.
Structural Biology Group, European Synchrotron Radiation Facility, Grenoble, France.
PLoS Biol. 2020 Nov 2;18(11):e3000936. doi: 10.1371/journal.pbio.3000936. eCollection 2020 Nov.
Using mRNA sequencing and de novo transcriptome assembly, we identified, cloned, and characterized 9 previously undiscovered fluorescent protein (FP) homologs from Aequorea victoria and a related Aequorea species, with most sequences highly divergent from A. victoria green fluorescent protein (avGFP). Among these FPs are the brightest green fluorescent protein (GFP) homolog yet characterized and a reversibly photochromic FP that responds to UV and blue light. Beyond green emitters, Aequorea species express purple- and blue-pigmented chromoproteins (CPs) with absorbances ranging from green to far-red, including 2 that are photoconvertible. X-ray crystallography revealed that Aequorea CPs contain a chemically novel chromophore with an unexpected crosslink to the main polypeptide chain. Because of the unique attributes of several of these newly discovered FPs, we expect that Aequorea will, once again, give rise to an entirely new generation of useful probes for bioimaging and biosensing.
利用 mRNA 测序和从头转录组组装,我们从维多利亚多管发光水母和一种相关的 Aequorea 物种中鉴定、克隆和表征了 9 种以前未发现的荧光蛋白(FP)同源物,其中大多数序列与维多利亚多管发光水母绿色荧光蛋白(avGFP)高度不同。这些 FP 中包括迄今为止所描述的最亮的绿色荧光蛋白(GFP)同源物和一种可逆光致变色 FP,它对 UV 和蓝光有响应。除了绿色发射器外,Aequorea 物种还表达具有从绿色到远红色的吸收范围的紫色和蓝色着色的色蛋白(CP),包括 2 种可光转化的 CP。X 射线晶体学揭示,Aequorea CP 含有一种具有意想不到的与主多肽链交联的化学新颖的生色团。由于这些新发现的 FP 中的几个具有独特的属性,我们预计 Aequorea 将再次产生新一代用于生物成像和生物传感的有用探针。