Expression and Crystallization of a Soluble Form of Drosophila Fasciclin III
A truncated form of Drosophila fasciclin III has bee engineered by site-directed mutagenesis. Secreted fasciclin III is expressed at 35 to 40 mg/l in insert cells with baculovirus carrying the recombinant gene. Single crystals of purified soluble fasciclin III have been grown by vapor diffusion versus polyethylene glycol 8000/sodium citrate at low pH. The space group is P6_122 or its enantiomorph P6_522, with unit cell dimensions ɑ = b = 140 Å, c = 260 Å. Cryo-preserved crystals diffract to reciprocal lattice spacing beyond 3·0 Å.
© 1994 Academic Press. This work was supported by a Basil O'Connor Starter Scholar Award (P.M.S.). the National Institute of Health (R29 NS28699 to P.M.S.) and the Howard Hughes Medical Institute (P.J.B.). P.J.B. was a Pew Scholar and held a Young Investigator award from the Cancer Research Institute during part of the work. R.K.S. was a post-doctoral fellow of the American Cancer Society, D.E.V. is supported by an NIH pre-doctoral training grant. We thank M.D. Summers for the gift of a baculovirus starter kit, Michael Blum and Leemor Joshua-Tor, who provided instruction on the cryo-preservation of maromolecular crystals, and Michael Soltis (SSRL.). Leemor Joshua-Tor and Michael Stowell for assistance with data collection at SSRL.