| Autor: |
Rueda S; Department of Chemistry, Brown University, Providence, Rhode Island 02912, United States., McCubbin TJ; Division of Plant Science and Technology, Interdisciplinary Plant Group, The Missouri Maize Center, University of Missouri, Columbia, Missouri 65211, United States., Shieh M; Department of Chemistry, Brown University, Providence, Rhode Island 02912, United States., Hoshing R; Department of Chemistry, Brown University, Providence, Rhode Island 02912, United States., Braun DM; Division of Plant Science and Technology, Interdisciplinary Plant Group, The Missouri Maize Center, University of Missouri, Columbia, Missouri 65211, United States.; Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211, United States., Basu A; Department of Chemistry, Brown University, Providence, Rhode Island 02912, United States. |
| Abstrakt: |
Small molecule fluorescent probes that bind selectively to plant cell wall polysaccharides have been instrumental in elucidating the localization and function of these glycans. Arabinogalactan proteins (AGPs) are cell wall proteoglycans implicated in essential functions such as cell signaling, plant growth, and programmed cell death. There is currently no small molecule probe capable of fluorescently labeling AGPs. The Yariv reagents are the only small molecules that bind AGPs, and have been used to study AGP function and isolate AGPs via precipitation of an AGP-Yariv complex. However, the Yariv reagents are not fluorescent, rendering them ineffective for localization studies using fluorescence microscopy. A fluorescent version of a Yariv reagent that is capable of both binding as well as imaging AGPs would provide a powerful tool for studying AGPs in planta. Herein, we describe the synthesis of an azido analog of the Yariv reagent that can be further functionalized with a fluorophore to provide a glycoconjugate that binds AGPs and is fluorescent. We show that the modified reagent binds gum arabic in in vitro binding assays when used in conjunction with the βGlcYariv reagent. Fluorescent imaging of AGPs in fixed maize leaf tissue enables localization of AGPs to cell walls in the leaf. Significantly, imaging can also be carried out using fresh tissue. This represents the first small molecule probe that can be used to visualize AGPs using fluorescence microscopy. |
