Expression of a long variant of CRACR2A that belongs to the Rab GTPase protein family in endothelial cells.
Bottom Line: Unexpectedly, short interfering RNA designed to deplete CRACR2A had no effect on CRAC channels in endothelial cells but reduced the abundance of a protein with about twice the mass of CRACR2A.It made a positive contribution to endothelial tube formation.The data suggest that endothelial cells contain a long variant of CRACR2A which is an EF-hand-containing Rab protein that lacks impact on CRAC channels.
Affiliation: School of Medicine, University of Leeds, Leeds LS2 9JT, UK.Show MeSH
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Mentions: An explanation for the above data could be that CRACR2A is not expressed. Indeed, anti-CRACR2A antibody failed to detect protein of ∼45 kDa (Fig. 2A), which is the expected mass of CRACR2A . However, a doublet around 95 kDa was labelled by the antibody and the lower of these two bands was depleted by two different siRNAs targeted to CRACR2A (Fig. 2A and B). The upper band may reflect an unrelated protein, labelled non-specifically by the antibody. The data suggest that CRACR2A is expressed in endothelial cells but that it occurs at about twice the expected molecular mass.
Affiliation: School of Medicine, University of Leeds, Leeds LS2 9JT, UK.