Function of a conserved sequence motif in biotin holoenzyme synthetases
- 1 January 2000
- journal article
- Published by Wiley in Protein Science
- Vol. 9 (8), 1530-1539
- https://doi.org/10.1110/ps.9.8.1530
Abstract
The biotin holoenzyme synthetases (BHS) are essential enzymes in all organisms that catalyze post-translational linkage of biotin to biotin-dependent carboxylases. The primary sequences of a large number of these enzymes are now available and homologies are found among all. The glycine-rich sequence, GRGRXG, constitutes one of the homologous regions in these enzymes and, based on its similarity to sequences found in a number of mononucleotide binding enzymes, has been proposed to function in ATP binding in the BHSs. In the Escherichia coli enzyme, the only member of the family for which a three-dimensional structure has been determined, the conserved sequence is found in a partially disordered surface loop. Mutations in the sequence have previously been isolated and characterized in vivo. In this work these single-site mutants, G115S, R118G, and R119W, of the E. coli BHS have been purified and biochemically characterized with respect to binding of small molecule substrates and the intermediate in the biotinylation reaction. Results of this characterization indicate that, rather than functioning in ATP binding, this glycine-rich sequence is required for binding the substrate biotin and the intermediate in the biotinylation reaction, biotinyl-5'-AMP. These results are of general significance for understanding structure-function relationships in biotin holoenzyme synthetases.Keywords
This publication has 42 references indexed in Scilit:
- Ligand-linked Structural Changes in the Escherichia coli Biotin Repressor: The Significance of Surface Loops for Binding and AllosteryJournal of Molecular Biology, 1999
- Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequenceNature, 1998
- Life with 6000 GenesScience, 1996
- MOLMOL: A program for display and analysis of macromolecular structuresJournal of Molecular Graphics, 1996
- Kinetics of Biotinyl-5'-adenylate Synthesis Catalyzed by the Escherichia coli Repressor of Biotin Biosynthesis and the Stability of the Enzyme-Product ComplexBiochemistry, 1994
- Crystal Structure of the DNA Binding Domain of the Heat Shock Transcription FactorScience, 1994
- Movable lobes and flexible loops in proteins Structural deformations that control biochemical activityFEBS Letters, 1993
- Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectorsGene, 1985
- Genetic and biochemical characterization of the birA gene and its product: Evidence for a direct role of biotin holoenzyme synthetase in repression of the biotin operon in Escherichia coliJournal of Molecular Biology, 1981
- The birA gene of Escherichia coli encodes a biotin holoenzyme synthetaseJournal of Molecular Biology, 1981