An atomic-resolution mechanism of 3-hydroxy-3-methylglutaryl–CoA synthase

Bahnson, Brian J.

doi:10.1073/pnas.0407418101

Cited by 11 publications

(9 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this mixed structure, each of the active site residues exhibits a single conformation indicating that this is an equilibrated state in which the phosphoryl group is being transferred. An equilibrated mixture of states in the enzyme reaction was also observed for 3-hydroxy-3-methylglutaryl-CoA synthase (25,26). For the phosphorylated BPGM, the covalent N-P bond length of phosphorylated His-11 is 1.7 Å, consistent with the calculated value of 1.76 Å (27).…”

Section: Structure Of Bpgm Bound With a 23-bpg Before Reaction-supporting

confidence: 75%

Seeing the Process of Histidine Phosphorylation in Human Bisphosphoglycerate Mutase

Wang

Liu

Wei

et al. 2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

Bisphosphoglycerate mutase is an erythrocyte-specific enzyme catalyzing a series of intermolecular phosphoryl group transfer reactions. Its main function is to synthesize 2,3-bisphosphoglycerate, the allosteric effector of hemoglobin. In this paper, we directly observed real-time motion of the enzyme active site and the substrate during phosphoryl transfer. A series of high resolution crystal structures of human bisphosphoglycerate mutase co-crystallized with 2,3-bisphosphoglycerate, representing different time points in the phosphoryl transfer reaction, were solved. These structures not only clarify the argument concerning the substrate binding mode for this enzyme family but also depict the entire process of the key histidine phosphorylation as a "slow movie". It was observed that the enzyme conformation continuously changed during the different states of the reaction. These results provide direct evidence for an "in line" phosphoryl transfer mechanism, and the roles of some key residues in the phosphoryl transfer process are identified.

show abstract

Section: Structure Of Bpgm Bound With a 23-bpg Before Reaction-supporting

confidence: 75%

Seeing the Process of Histidine Phosphorylation in Human Bisphosphoglycerate Mutase

Wang

Liu

Wei

et al. 2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…After dissociation of CoASH, the second substrate acetoacetyl‐CoA enters the catalytic site, forming the HMG‐CoA–enzyme complex. In the final step, HMG‐CoA is produced by the hydrolysis of HMG‐CoA–enzyme complex (Bahnson, ). The binary structure of acetoacetyl‐CoA explains the covalent bond between reduced CoA and acetoacetate at the active site cysteine.…”

Section: Potential Drug Targetssupporting

confidence: 67%

In search of novel protein drug targets for treatment of Enterococcus faecalis infections

et al. 2019

View full text Add to dashboard Cite

Enterococcus faecalis (Ef) is one of the major pathogens involved in hospital-acquired infections. It can cause nosocomial bacteremia, surgical wound infection, and urinary tract infection. It is important to mention here that Ef is developing resistance against many commonly occurring antibiotics. The occurrence of multidrug resistance (MDR) and extensive-drug resistance (XDR) is now posing a major challenge to the medical community. In this regard, to combat the infections caused by Ef, we have to look for an alternative. Rational structure-based drug design exploits the three-dimensional structure of the target protein, which can be unraveled by various techniques such as X-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. In this review, we have discussed the complete picture of Ef infections, the possible treatment available at present, and the alternative treatment options to be explored. This study will help in better understanding of novel biological targets against Ef and the compounds, which are likely to bind with these targets. Using these detailed structural informations, rational structure-based drug design is achievable and tight inhibitors against Ef can be prepared. K E Y W O R D SEnterococcus faecalis, multidrug-resistant bacteria, protein drug targets, structure-based drug design

show abstract

“…A small amount of nonenzymatic loading of holo-PksL by Ac-CoA was also observed by autoradiography and mass spectrometry (data not shown). When the experiment was repeated in the absence of PksL-T2, the radiolabel accumulated on PksG, suggesting that the reaction proceeds by an Ac-PksG intermediate, consistent with the mechanism of HMG-CoA synthases (22) (Fig. 12, which is published as supporting information on the PNAS web site).…”

Section: Resultsmentioning

confidence: 59%

“…We hypothesized that PksG catalyzed the transfer of OCH 2 COO Ϫ from Ac-S-AcpK to a ␤-ketothioester polyketide intermediate linked to one of the consecutive thiolation domains of PksL, in a reaction analogous to that catalyzed by HMG-CoA synthase (22,23). We chose acetoacetyl (Acac)-S-PksL-T2 as a model substrate for this reaction.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Convergence of isoprene and polyketide biosynthetic machinery: Isoprenyl- S -carrier proteins in the pksX pathway of Bacillus subtilis

Calderone

Kowtoniuk

Kelleher

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

169

230

View full text Add to dashboard Cite

The pksX gene cluster from Bacillus subtilis is predicted to encode the biosynthesis of an as yet uncharacterized hybrid nonribosomal peptide͞polyketide secondary metabolite. We used a combination of biochemical and mass spectrometric techniques to assign functional roles to the proteins AcpK, PksC, PksL, PksF, PksG, PksH, and PksI, and we conclude that they act to incorporate an acetate-derived ␤-methyl branch on an acetoacetyl-S-carrier protein and ultimately generate a ⌬ 2 -isoprenyl-S-carrier protein. This work highlights the power of mass spectrometry to elucidate the functions of orphan biosynthetic enzymes, and it details a mechanism by which single-carbon ␤-branches can be inserted into polyketide-like structures. This pathway represents a noncanonical route to the construction of prenyl units and serves as a prototype for the intersection of isoprenoid and polyketide biosynthetic manifolds in other natural product biosynthetic pathways.mass spectrometry ͉ orphan gene cluster ͉ hybrid nonribosomal peptide͞polyketide ͉ polyketide methylation P olyketides and nonribosomal peptides are classes of secondary metabolites that are synthesized by the iterative coupling of malonyl (Mal) derivatives and amino acids, respectively. The catalytic machinery responsible for the biosynthesis of these natural products comprises modular synthases that incorporate, and often subsequently tailor, monomer units into the growing small molecule by a thiotemplated mechanism. Often, the sequence of protein domains and modules is colinear with respect to the sequence of biosynthetic reactions catalyzed by the polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) machinery (1-3). Because of their highly modular nature, the small-molecule products of PKSs, NRPSs, and hybrid NRPS-PKS systems can often be predicted by bioinformatic approaches, and there have been recent experimental validations of such predictions (4-6). However, for many cases in which the biosynthetic product encoded by a given gene cluster is unknown, functional characterization of the synthase by bioinformatic methods alone is difficult or impossible.One such orphan cluster for which the product structure is unknown is the hybrid NRPS-PKS pksX cluster from Bacillus subtilis (7-9). Although the pksX cluster has been proposed to encode the biosynthesis of difficidin (7,9,10), recent work has demonstrated that the NRPS portion of the gene cluster is active, disqualifying difficidin, which does not contain any amines, as a candidate for the biosynthetic product of this cluster (11). Long thought to be cryptic, there is now evidence that it is responsible for the biosynthesis of a product that kills streptomycetes (P. D. Straight, M. A. Fischbach, C.T.W., and R. Kolter, unpublished results). Additionally, this cluster does not follow the usual NRPS-PKS colinearity rules because it has only three trans-acting acyltransferases that may load up to 17 of the 20 predicted thiolation domains of the cluster.We were intrigued by a series of predicted ORFs withi...

show abstract

An atomic-resolution mechanism of 3-hydroxy-3-methylglutaryl–CoA synthase

Cited by 11 publications

References 19 publications

Seeing the Process of Histidine Phosphorylation in Human Bisphosphoglycerate Mutase

Seeing the Process of Histidine Phosphorylation in Human Bisphosphoglycerate Mutase

In search of novel protein drug targets for treatment of Enterococcus faecalis infections

Convergence of isoprene and polyketide biosynthetic machinery: Isoprenyl- S -carrier proteins in the pksX pathway of Bacillus subtilis

Contact Info

Product

Resources

About