An automated microseed matrix-screening method for protein crystallization.
A microseed-matrix procedure has been established with the aim of influencing the nucleation event in standard crystallization screens. The method is based on the original description of matrix seeding described by Ireton & Stoddard (2004, Acta Cryst. D60, 601-605). Seed stocks are produced using a simple "seed-bead" method. The protein, reservoir solutions and seed stocks are pipetted simultaneously using a three-bore dispensing tip in drops of 0.6 microl total volume. The number and type of hits produced with the proteins tested in this study has been increased and it is believed that this method could be generally applicable to proteins where little or no nucleation is normally observed.
D'Arcy
Allan
A
Novartis Institutes of Biomedical Research, Protease Platform, Klybeckstrasse 144, CH 4002 Basel, Switzerland. allan.darcy@novartis.com
Villard
Frederic
F
Marsh
May
M
D000818:Animals; D001331:Automation; D002417:Cattle; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D006801:Humans; D053508:Matrix Metalloproteinase 12 / Q000737:chemistry; D010196:Pancreatic Elastase / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D013552:Swine; D014357:Trypsin / Q000737:chemistry; D043222:Ubiquitin Thiolesterase / Q000737:chemistry; D000076084:Ubiquitin-Specific Peptidase 7
D011506:Proteins; C464569:USP7 protein, human; D043222:Ubiquitin Thiolesterase; D000076084:Ubiquitin-Specific Peptidase 7; D010196:Pancreatic Elastase; D014357:Trypsin; D053508:Matrix Metalloproteinase 12