By James W Putney
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Extra info for Calcium signaling
An interesting offshoot of the fluo-4 work is the development of fluo-4 analogs linked to biotin and streptavidin as well as amine-reactive derivatives. The synthesis of these dextrans aims for one molecule of indicator per molecule of dextran but in practice the ratio can vary. Since the indicator-to-dextran ratio can affect the affinity, Kd needs to be determined for each batch [26, 28]. The size (MW) of dextran should also be considered since entry into the nucleus can vary with molecular weight [96, 97].
When calcium measurements were made using fura-2, the differences in kinetics and magnitude of the calcium transient were immediately evident. Attempts to resolve this disparity continue but at least two factors have emerged. One of these factors concerns the intrinsic properties of the various indicators especially with respect to their on- and off-rates for calcium binding, their dissociation constants, and their ability to buffer calcium inside the cell. Another factor concerns interactions between the indicator and its intracellular environment.
Since the introduction of furaptra, a wide variety of low-affinity tricarboxylate indicators have been developed by Molecular Probes. These are all tricarboxylate counterparts of the BAPTA-based calcium indicators. Generally, the optical properties of the tricarboxylate indicators are identical to that of the tetracarboxylate counterparts. The key differences between the various furaptra analogs lie in their James W. 10 Chemical structures of low-affinity calcium indicators. For the fluorinated indicators F ¼ fluorine and H ¼ hydrogen.
Calcium signaling by James W Putney