Abstract
Nitric oxide synthase like enzyme (NOS-like enzyme), which produces nitric oxide, participates in many biological processes. However it remains unidentified and highly controversial that plants do possess a NOS-like enzyme. In this paper, a novel arginine analogue NP1 was designed and developed for the direct identification and real time tracking of NOS-like enzymes in plant by fluorescence sensing. It could bind NOS-like enzyme efficiently and enter the cell successfully. In vivo fluorescence response results directly proved that NOS-like enzymes did exist in tobacco leaf and would be stimulated by pathogen infection, which also provided a useful chemical tool for the study of the function of NOS-like enzyme in plants.
Highlights
Antibodies against mammalian NOS may recognize many other proteins not connected with NOS
Eric Deprez et al have reported a two-photon fluorescent sensor which was used for the biological imaging of eNOS in living human umbilical vein endothelial cells[23]
There were much fewer reports about fluorescent sensors used in plants, one reason is that the existence of the plant cell wall, small molecule fluorescent sensor is difficult to enter the plant cell
Summary
Kang Chang1,*, Tongtong Guo2,*, Pengfei Li1, Yin Liu[2], Yufang Xu1, Yuda Fang2 & Xuhong Qian[1]. A novel arginine analogue NP1 was designed and developed for the direct identification and real time tracking of NOS-like enzymes in plant by fluorescence sensing It could bind NOS-like enzyme efficiently and enter the cell successfully. Upon the receptor of L-NAME combined with NOS-like enzyme, the conformation of piperazine would change from “boat form” to “chair form”, leading to the nitrogen atom away from the xanthene ring, photo-induced electron transfer (PET) effect was quenched and the fluorescence would be recovered With this fluorescent arginine analogue NP1 in hand, we can realize the direct identification and tracking of NOS-like enzyme in plants by fluorescence sensing using the mechanism of conformational photo-induced electron transfer (PET)[10,14,30]
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