title: Controlled in meso phase crystallization--a method for the structural investigation of membrane proteins.
abstract: We investigated in meso crystallization of membrane proteins to develop a fast screening technology which combines features of the well established classical vapor diffusion experiment with the batch meso phase crystallization, but without premixing of protein and monoolein. It inherits the advantages of both methods, namely (i) the stabilization of membrane proteins in the meso phase, (ii) the control of hydration level and additive concentration by vapor diffusion. The new technology (iii) significantly simplifies in meso crystallization experiments and allows the use of standard liquid handling robots suitable for 96 well formats. CIMP crystallization furthermore allows (iv) direct monitoring of phase transformation and crystallization events. Bacteriorhodopsin (BR) crystals of high quality and diffraction up to 1.3 Å resolution have been obtained in this approach. CIMP and the developed consumables and protocols have been successfully applied to obtain crystals of sensory rhodopsin II (SRII) from Halobacterium salinarum for the first time.
authors.affiliation: QIAGEN GmbH, Hilden, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Kubicek
authors.forename: Ramona
authors.initials: R
authors.lastname: Schlesinger
authors.forename: Christian
authors.initials: C
authors.lastname: Baeken
authors.forename: Georg
authors.initials: G
authors.lastname: Büldt
authors.forename: Frank
authors.initials: F
authors.lastname: Schäfer
authors.forename: Jörg
authors.initials: J
authors.lastname: Labahn
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D005989:Glycerides / Q000737:chemistry; D019617:Halobacterium salinarum; D025602:Halorhodopsins / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D044367:Phase Transition; D055550:Protein Stability; D025603:Sensory Rhodopsins / Q000737:chemistry*
chemical_list: D005989:Glycerides; D025602:Halorhodopsins; D008565:Membrane Proteins; D025603:Sensory Rhodopsins; C061508:sensory rhodopsin II protein, archaeal; D001436:Bacteriorhodopsins; C005953:monoolein