Methods in molecular biology (Clifton, N.J.)

Longistatin, an EF-hand Ca2+-binding protein from vector tick: identification, purification, and characterization.

PMID 23296609


EF-hand Ca(2+)-binding motif, a structural component of the EF-hand protein, functions as a calcium sensor and/or buffer in the cytosol of the cell. However, in a few exceptional cases, the EF-hand proteins are secreted from cells and play crucial roles extracellularly. We have identified longistatin, an EF-hand Ca(2+)-binding protein, from the salivary glands of the tick, Haemaphysalis longicornis. Longistatin possesses an N-terminal sequence of unknown structure and two EF-hand motifs in the C-terminus, which conserve a calmodulin-like canonical structure. Longistatin shows distinct changes in its migration during electrophoresis through SDS-PAGE gel containing calcium or ethylenediaminetetraacetic acid (EDTA). Both recombinant and endogenous forms of longistatin can be stained with rutheninum red, demonstrating that longistatin is a Ca(2+)-binding protein.