Calretinin--sensor or buffer--function still unclear.

Calretinin (CR) is a 31.5 kDa EF-hand calcium binding protein. CR has a general neuronal localization and is a predominantly cytosolic protein. The biochemical properties of this protein are well characterized but its function is still unclear. It is often postulated that the presence of CR correlates with an increased survival ability of cells under pathological conditions connected with increased intracellular calcium levels. However, not all studies confirm such a relationship and they indicate that the presence of CR does not protect cells against calcium overload and that CR does not act as an intracellular calcium buffer. Recent results concerning CR function are discussed and the conclusion is proposed that CR acts as a calcium modulator rather than a calcium buffer.

Measurements of [Ca2+] using fura-2 in glioma C6 cells expressing calretinin with GFP as a marker of transfection: no Ca2+-buffering provided by calretinin.

Glioma C6 cells were transfected with a plasmid containing the calretinin (CR) and green fluorescent protein (GFP) coding regions to analyze the effect of CR's presence on [Ca2+]i. Positive transfectants were identified by the detection of GFP and [Ca2+]i was measured using fura-2 as a probe. We found that neither the basic [Ca2+]i nor activated [Ca2+]i achieved by exposure to ionomycin, ADP or thapsigargin were affected by CR's presence in transfected cells, despite the ability of CR to bind Ca2+ as part of fusion protein. The level of expressed CR was estimated as at least 1 microM. The presented results suggest that CR's function is unlikely to be an intracellular Ca2+-buffer and support the hypothesis that CR might be involved in a specific Ca2+-dependent process. The results of this work also show that the S65T mutant of GFP is compatible with fura-2 measurements of intracellular [Ca2+]. We have demonstrated that the presence of GFP, as a transfection marker of glioma C6 cells, does not disturb fura-2 fluorescence, the basal or activated [Ca2+]i in these cells.