Synthesis of a BODIPY library and its application to the development of live cell glucagon imaging probe.
The first BODIPY library (BD) was synthesized, and a highly selective glucagon sensor, Glucagon Yellow (BD-105), was discovered by fluorescence image-based screening method. BD library was synthesized via a Knoevenagel-type condensation reaction with 160 benzaldehydes and the 1,3 dimethyl-BODIPY scaffold. Using BD compounds, a fluorescence image-based screening was performed against three cell lines including AlphaTC1 and BetaTC6 cells which secret glucagon and insulin, respectively, and HeLa as control cells. Out of the 160 candidate probes, one compound, Glucagon Yellow, exhibited selective staining only in AlphaTC1 cells. The selectivity of Glucagon Yellow toward glucagon was confirmed in vitro by comparison of its fluorescence intensity change against 19 biologically relevant analytes. Subsequent immunostaining experiments revealed that Glucagon Yellow and the glucagon antibody colocalized in pancreas tissue, showing a high quantitative correlation analysis by the Pearson's coefficient constant (R(r) = 0.950). These results demonstrated the potential application of Glucagon Yellow as a glucagon imaging agent in live cells and tissues.
Lee
Jun-Seok
JS
Department of Chemistry, New York University, New York, New York 10003, USA.
Kang
Nam-young
NY
Kim
Yun Kyung
YK
Samanta
Animesh
A
Feng
Suihan
S
Kim
Hyeong Kyu
HK
Vendrell
Marc
M
Park
Jung Hwan
JH
Chang
Young-Tae
YT
D041721:3T3-L1 Cells; D000818:Animals; D001896:Boron Compounds / Q000032:analysis* / Q000138:chemical synthesis* / Q000737:chemistry; D002460:Cell Line; D002470:Cell Survival; D005456:Fluorescent Dyes / Q000032:analysis / Q000138:chemical synthesis* / Q000737:chemistry; D005934:Glucagon / Q000031:analogs & derivatives / Q000032:analysis* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D015394:Molecular Structure; D011789:Quantum Theory; D054852:Small Molecule Libraries
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D054852:Small Molecule Libraries; C544198:glucagon yellow; D005934:Glucagon