Crystallization and Preliminary X-ray Crystallographic Studies of the Alkanesulfonate FMN Reductase from Escherichia coli

Journal Article · Sat Jan 01 04:00:00 EST 2005 · Acta Cryst. F

DOI:https://doi.org/10.1107/S1744309105024206· OSTI ID:913889

Gao, B; Bertrand, A; Boles, W; Ellis, H; Mallett, T

The alkanesulfonate FMN reductase (SsuE) from Escherichia coli catalyzes the reduction of FMN by NADPH to provide reduced flavin for the monooxygenase (SsuD) enzyme. The vapor-diffusion technique yielded single crystals that grow as hexagonal rods and diffract to 2.9 Angstrom resolution using synchrotron X-ray radiation. The protein crystallizes in the primitive hexagonal space group P622. The SsuE protein lacks any cysteine or methionine residues owing to the role of the SsuE enzyme in the acquisition of sulfur during sulfate starvation. Therefore, substitution of two leucine residues (Leu114 and Leu165) to methionine was performed to obtain selenomethionine-containing SsuE for MAD phasing. The selenomethionine derivative of SsuE has been expressed and purified and crystals of the protein have been obtained with and without bound FMN. These preliminary studies should lead to the structure solution of SsuE. It is anticipated that this new protein structure will provide detailed structural information on specific active-site regions of the protein and insight into the mechanism of flavin reduction and transfer of reduced flavin.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 913889
• Report Number(s): BNL--78457-2007-JA
• Journal Information: Acta Cryst. F, Journal Name: Acta Cryst. F Vol. 61
• Country of Publication: United States
• Language: English

Related Subjects

43 PARTICLE ACCELERATORS
CRYSTALLIZATION
CRYSTALLOGRAPHY
ESCHERICHIA COLI
MONOCRYSTALS
PROTEIN STRUCTURE
SPACE GROUPS
SYNCHROTRON RADIATION
national synchrotron light source