title: NMR studies on the 46-kDa dimeric protein, 3,4-dihydroxy-2-butanone 4-phosphate synthase, using 2H, 13C, and 15N-labelling.
abstract: 3,4-Dihydroxy-2-butanone 4-phosphate synthase catalyses the release of C-4 from the substrate, ribulose phosphate, via a complex series of rearrangement reactions. The cognate ribB gene of Escherichia coli was hyperexpressed in a recombinant E. coli strain. The protein was shown to be a 46-kDa homodimer by hydrodynamic analysis. A variety of protein samples labelled with different grades of 13C, 15N and 2H, i.e. one with 100% 2H and 15N, one with 75% 2H, 99% 13C, 15N, and one with 100% 2H, 99% 13C,15N were prepared. Despite the large molecular size, 2- and 3-dimensional NMR spectra of reasonable quality were obtained. Attempts at the assignment of individual 13C, 15N and 1H signals show, in principle, the feasibility of structure determination. The number of NMR signals shows unequivocally that the homodimeric protein obeys strict C2 symmetry.
authors: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Garching, Germany.
authors: G
authors: G
authors: Richter
authors: M
authors: M
authors: Kelly
authors: C
authors: C
authors: Krieger
authors: Y
authors: Y
authors: Yu
authors: W
authors: W
authors: Bermel
authors: G
authors: G
authors: Karlsson
authors: A
authors: A
authors: Bacher
authors: H
authors: H
authors: Oschkinat
mesh_terms: D001483:Base Sequence; D002247:Carbon Isotopes; D017931:DNA Primers / Q000235:genetics; D003903:Deuterium; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D009587:Nitrogen Isotopes
chemical_list: D002247:Carbon Isotopes; D017931:DNA Primers; D009587:Nitrogen Isotopes; D003903:Deuterium; D019751:Intramolecular Transferases; C066493:L-3,4-dihydroxy-2-butanone-4-phosphate synthase