Pham Elizabeth, Mills Evan, Truong Kevin
Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, ON M5S 3G9, Canada.
Chem Biol. 2011 Jul 29;18(7):880-90. doi: 10.1016/j.chembiol.2011.04.014.
Ca(2+) signals regulate diverse physiological processes through tightly regulated fluxes varying in location, time, frequency, and amplitude. Here, we developed LOVS1K, a genetically encoded and photoactivated synthetic protein to generate local or global Ca(2+) signals. With 300 ms blue light exposure, LOVS1K translocated to Orai1, a plasma membrane Ca(2+) channel, within seconds, generating a local Ca(2+) signal on the plasma membrane, and returning to the cytoplasm after tens of seconds. With repeated photoactivation, global Ca(2+) signals in the cytoplasm were generated to modulate engineered Ca(2+)-inducible proteins. Although Orai1 is typically associated with global store-operated Ca(2+) entry, we demonstrate that Orai1 can also generate local Ca(2+) influx on the plasma membrane. Our photoactivation system can be used to generate spatially and temporally precise Ca(2+) signals and to engineer synthetic proteins that respond to specific Ca(2+) signals.
钙离子信号通过在位置、时间、频率和幅度上严格调控的通量来调节多种生理过程。在此,我们开发了LOVS1K,一种基因编码的光激活合成蛋白,用于产生局部或全局钙离子信号。在300毫秒蓝光照射下,LOVS1K在数秒内转运至质膜钙离子通道Orai1,在质膜上产生局部钙离子信号,并在数十秒后返回细胞质。通过重复光激活,可在细胞质中产生全局钙离子信号以调节工程化的钙离子诱导蛋白。尽管Orai1通常与全局储存-操作性钙离子内流相关,但我们证明Orai1也可在质膜上产生局部钙离子内流。我们的光激活系统可用于产生时空精确的钙离子信号,并设计对特定钙离子信号作出反应的合成蛋白。