Fluorescent indicators including a binding protein moiety, a donor
fluorescent protein moiety, and an acceptor fluorescent protein moiety
are described. The binding protein moiety has an analyte-binding region
which binds an analyte and causes the indicator to change conformation
upon exposure to the analyte. The donor moiety and the acceptor moiety
change position relative to each other when the analyte binds to the
analyte-binding region. The donor moiety and the acceptor moiety exhibit
fluorescence resonance energy transfer when the donor moiety is excited
and the distance between the donor moiety and the acceptor moiety is
small. The indicators can be used to measure analyte concentrations in
samples, such as calcium ion concentrations in cells.