title: Protein products obtained by site-preferred partial crosslinking in protein crystals and "liberated" by redissolution.
abstract: The use of protein crystals as a source of nanoscale biotemplates has attracted growing interest in recent years owing to their inherent internal order. As these crystals are vulnerable to environmental changes, potential applications require their stabilization by chemical crosslinking. We have previously shown that such intermolecular chemical crosslinking reactions occurring within protein crystals are not random events, but start at preferred crosslinking sites imposed by the alignment of protein molecules and their packing within the crystalline lattice. Here we propose a new working hypothesis and demonstrate its feasibility in enabling us to extricate homogeneous populations of single protein molecules that display chemical point mutations or of dimers that show homogeneous chemical crosslinking, and that have the potential for isolation of higher structures. Characterization of the crosslinking mechanism and its end products opens the way to the potential retrieval of such specific modified/intermolecular crosslinked products simply by effecting partial crosslinking at identified preferred sites, followed by time-controlled arrest of the crosslinking reaction and dissolution of the crystals by medium exchange complemented by chromatographic purification.
authors.affiliation: Department of Molecular Microbiology and Biotechnology, Tel Aviv University, 69978, Tel-Aviv, Israel.
authors.forename: Michal
authors.initials: M
authors.lastname: Buch
authors.forename: Yariv
authors.initials: Y
authors.lastname: Wine
authors.forename: Yael
authors.initials: Y
authors.lastname: Dror
authors.forename: Sonia
authors.initials: S
authors.lastname: Rosenheck
authors.forename: Mario
authors.initials: M
authors.lastname: Lebendiker
authors.forename: Rita
authors.initials: R
authors.lastname: Giordano
authors.forename: Ricardo M F
authors.initials: RM
authors.lastname: Leal
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Popov
authors.forename: Amihay
authors.initials: A
authors.lastname: Freeman
authors.forename: Felix
authors.initials: F
authors.lastname: Frolow
mesh_terms: D003460:Crystallization; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins
keywords: X-ray diffraction; crosslinking; glutaraldehyde; protein crystals; structure analysis