title: Purification, crystallization and preliminary X-ray diffraction analysis of the lytic transglycosylase MltF from Escherichia coli.
abstract: The lytic transglycosylase MltF from Escherichia coli is an outer-membrane-bound periplasmic protein with two domains: a C-terminal catalytic domain with a lysozyme-like fold and an N-terminal domain of unknown function that is homologous to the periplasmic substrate-binding proteins of ABC transporters. In order to investigate its structure and function, a soluble form of full-length MltF (sMltF) containing both domains and a soluble fragment containing only the N-terminal domain (sMltF-NTD) were purified and crystallized. Crystals of sMltF belonged to space group P4(3)2(1)2 or P4(1)2(1)2, with unit-cell parameters a = b = 110.8, c = 163.5 A and one or two molecules per asymmetric unit. A complete data set was collected to 3.5 A resolution. Crystals of sMltF-NTD belonged to space group P3(1)21, with unit-cell parameters a = b = 82.4, c = 75.2 A and one molecule per asymmetric unit. For sMltF-NTD, a complete native data set was collected to 2.20 A resolution. In addition, for phasing purposes, a three-wavelength MAD data set was collected to 2.5 A resolution using a bromide-soaked sMltF-NTD crystal. Using phases derived from the Br-MAD data, it was possible to build a partial model of sMltF-NTD.
authors.affiliation: Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Pramod K
authors.initials: PK
authors.lastname: Madoori
authors.forename: Andy Mark W H
authors.initials: AM
authors.lastname: Thunnissen
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification; D016695:Glycosyltransferases / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D029968:Escherichia coli Proteins; D016695:Glycosyltransferases; C528867:MltF protein, E coli
references.citation: Nature. 1994 Feb 24;367(6465):750-3
references.pmid: 8107871
references.citation: Proteins. 1995 Jul;22(3):245-58
references.pmid: 7479697
references.citation: Int J Biochem Cell Biol. 2008;40(4):586-91
references.pmid: 17468031
references.citation: J Mol Biol. 1999 Aug 27;291(4):877-98
references.pmid: 10452894
references.citation: J Mol Evol. 2001 Jan;52(1):78-84
references.pmid: 11139297
references.citation: Microbiol Mol Biol Rev. 1998 Mar;62(1):181-203
references.pmid: 9529891
references.citation: J Mol Biol. 2005 Oct 7;352(5):1068-80
references.pmid: 16139297
references.citation: FEMS Microbiol Rev. 2008 Mar;32(2):149-67
references.pmid: 18194336
references.citation: Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1174-82
references.pmid: 12832760
references.citation: Acta Crystallogr D Biol Crystallogr. 1996 Jan 1;52(Pt 1):30-42
references.pmid: 15299723
references.citation: Acta Crystallogr D Biol Crystallogr. 2000 Feb;56(Pt 2):232-7
references.pmid: 10666615
references.citation: Microbiol Rev. 1993 Jun;57(2):320-46
references.pmid: 8336670
references.citation: Structure. 1999 Oct 15;7(10):1167-80
references.pmid: 10545329
references.citation: J Biol Chem. 2008 Mar 28;283(13):8363-73
references.pmid: 18234673
references.citation: J Bacteriol. 2002 Nov;184(22):6093-9
references.pmid: 12399477
references.citation: EXS. 1996;75:425-9
references.pmid: 8765311
references.citation: J Biol Chem. 2007 Jul 20;282(29):21197-205
references.pmid: 17502382
references.citation: Cell Mol Life Sci. 2003 Nov;60(11):2371-88
references.pmid: 14625683
references.citation: Biochemistry. 2000 Feb 29;39(8):1924-34
references.pmid: 10684641
references.citation: Methods Mol Biol. 2007;364:215-30
references.pmid: 17172768
references.citation: Microbiol Mol Biol Rev. 2008 Jun;72(2):211-27, table of contents
references.pmid: 18535144
references.citation: J Bacteriol. 1998 Jul;180(13):3441-7
references.pmid: 9642199
references.citation: J Am Chem Soc. 2009 Jul 1;131(25):8742-3
references.pmid: 19496566