Purification, crystallization and X-ray diffraction analysis of human synaptotagmin 1 C2A-C2B.

ABSTRACT

Synaptotagmin acts as the Ca(2+) sensor for neuronal exocytosis. The cytosolic domain of human synaptotagmin 1 is composed of tandem C2 domains C2A and C2B. These C2 domains modulate the interaction of synaptotagmin with the phospholipid bilayer of the presynaptic terminus and effector proteins such as the SNARE complex. Human synaptotagmin C2A-C2B has been expressed as a glutathione-S-transferase fusion protein in Escherichia coli. The purification, crystallization and preliminary X-ray analysis of this protein are reported here. The crystals diffract to 2.7 A and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 82.37, b = 86.31, c = 140.2 A. From self-rotation function analysis, there are two molecules in the asymmetric unit. The structure determination of the protein using this data is ongoing.