title: Crystallization and preliminary analysis of native and N-terminal truncated isoforms of toluene-4-monooxygenase catalytic effector protein.
abstract: Single crystals have been obtained of the toluene 4-monooxygenase catalytic effector protein, the SeMet-enriched protein and a truncated isoform missing ten amino acids from the N-terminus. Complete X-ray diffraction data sets have been collected and analyzed to 2.0, 3.0 and 1.96 A resolution for the native, SeMet and truncated isoform crystals, respectively. The native and SeMet proteins crystallized in space group P6(1)22 (unit-cell parameters a = b = 86.41 +/- 0.15, c = 143.90 +/- 0.27 A), whereas the truncated isoform crystallized in space group P2(1)3 (a = b = c = 86.70 +/- 0.47 A). Matthews coefficient calculations suggest either two or three molecules per asymmetric unit in the P6(1)22 space group and two molecules per asymmetric unit in the P2(1)3 space group. Experimental phases from MAD analysis of the SeMet isoform and molecular replacement of the truncated isoform confirm the presence of two molecules per asymmetric unit in each case. These crystallographic results are the first available for the evolutionarily related but functionally diversified catalytic effector proteins from the multicomponent diiron monooxygenase family.
authors.affiliation: School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332-0400, USA. allen.orville@chemistry.gatech.edu
authors.forename: Allen M
authors.initials: AM
authors.lastname: Orville
authors.forename: Joey M
authors.initials: JM
authors.lastname: Studts
authors.forename: George T
authors.initials: GT
authors.lastname: Lountos
authors.forename: Kevin H
authors.initials: KH
authors.lastname: Mitchell
authors.forename: Brian G
authors.initials: BG
authors.lastname: Fox
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D010105:Oxygenases / Q000737:chemistry* / Q000302:isolation & purification; D011549:Pseudomonas / Q000201:enzymology
chemical_list: D007527:Isoenzymes; D010105:Oxygenases; C068610:toluene-4-monooxygenase