title: Crystallization and preliminary X-ray crystallographic studies of the biotin carboxyl carrier protein and biotin protein ligase complex from Pyrococcus horikoshii OT3.
abstract: Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. To elucidate the exact details of the protein-protein interactions in the biotinylation function, the C-terminal half fragment of BCCP (BCCPDeltaN76), the R48A mutant of BPL (BPL*) and the R48A K111A double mutant of BPL (BPL**), all of which are from Pyrococcus horikoshii OT3, have been expressed, purified and successfully cocrystallized. Cocrystals of the BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 complexes as well as crystals of BPL*, BPL** and BCCPDeltaN76 were obtained by the oil-microbatch method using PEG 20 000 as a precipitant at 295 K. Complete X-ray diffraction data sets for BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 crystals were collected at 100 K to 2.7 and 2.0 A resolution, respectively, using synchrotron radiation. They belong to the monoclinic space group P2(1), with similar unit-cell parameters a = 69.85, b = 63.12, c = 75.64 A, beta = 95.9 degrees . Assuming two subunits of the complex per asymmetric unit gives a V(M) value of 2.45 A(3) Da(-1) and a solvent content of 50%.
authors.affiliation: Advanced Protein Crystallography Research Group, RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan.
authors.forename: Bagautdin
authors.initials: B
authors.lastname: Bagautdinov
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Matsuura
authors.forename: Svetlana
authors.initials: S
authors.lastname: Bagautdinova
authors.forename: Naoki
authors.initials: N
authors.lastname: Kunishima
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000737:chemistry* / Q000235:genetics; D019731:Carbon-Nitrogen Ligases / Q000737:chemistry* / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D002853:Chromatography, Liquid; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D054889:Fatty Acid Synthase, Type II; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D043902:Pyrococcus horikoshii / Q000201:enzymology*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D002352:Carrier Proteins; D054889:Fatty Acid Synthase, Type II; D019731:Carbon-Nitrogen Ligases; D000103:Acetyl-CoA Carboxylase; C079633:biotin carboxyl carrier protein