Biochemical and Structural Study of the Atypical Acyltransferase Domain from the Mycobacterial Polyketide Synthase Pks13

Bergeret, Fabien; Gavalda, Sabine; Chalut, Christian; Malaga, Wladimir; Quémard, Annaık̈; Pédelacq, J.D.; Daffé, Mamadou; Guilhot, Christophe; Mourey, Lionel; Bon, Cécile

doi:10.1074/jbc.m111.325639

Cited by 44 publications

(53 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The linker-AT unit appears to be a fixed unit in cis -AT PKS pathways. For example, guided by the domain architecture from the original KS-AT di-domain structures, two groups excised linker-AT units and obtained crystal structures [47,48] in which the interfaces of KS/AT linker domain and AT domain are identical to those of the PikAIII full module (Figure 3c). In contrast to the KS/AT linker domain, a small ferredoxin-like sub-domain of the AT occupies more varied positions relative to the AT core.…”

Section: Overall Architecturementioning

confidence: 99%

Architecture of the polyketide synthase module: surprises from electron cryo-microscopy

Smith

Skiniotis

Sherman

2015

Current Opinion in Structural Biology

View full text Add to dashboard Cite

Modular polyketide synthases produce a vast array of bioactive molecules that are the basis of many highly valued pharmaceuticals. The biosynthesis of these compounds is based on ordered assembly lines of multi-domain modules, each extending and modifying a specific chain-elongation intermediate before transfer to the next module for further processing. The first 3D structures of a full polyketide synthase module in different functional states were obtained recently by electron cryo-microscopy. The unexpected module architecture revealed a striking evolutionary divergence of the polyketide synthase compared to its metazoan fatty acid synthase homolog, as well as remarkable conformational rearrangements dependent on its biochemical state during the full catalytic cycle. The design and dynamics of the module are highly optimized for both catalysis and fidelity in the construction of complex, biologically active natural products.

show abstract

Section: Overall Architecturementioning

confidence: 99%

Architecture of the polyketide synthase module: surprises from electron cryo-microscopy

Smith

Skiniotis

Sherman

2015

Current Opinion in Structural Biology

View full text Add to dashboard Cite

show abstract

“…Biocatalytic chemistries centered on the amide bond are of fundamental industrial importance [45] and have widespread applications ranging from fine chemical synthesis to the generation of biopharmaceuticals [46]. By focusing on the α/β-phospholipase fold that forms an integral part of binding proteins [47] and that is furthermore capable of harnessing a plethora of activities including acyl transfer in polyketide biosynthetic machineries [48], a previously unknown enzyme-assisted hydrogen bond acceptor for stabilizing nitrogen inversion was discovered (Figure 2). Whereas the molecular mechanisms behind previously reported promiscuous amidase activities in esterase scaffolds (see Table 1 for some examples) are not completely understood or are suggested to depend upon water-mediated interactions [39,49] whose evolutionary role remains unclear [29], the present study reports on the first example of a direct enzyme-assisted hydrogen bond acceptor.…”

Section: Discussionmentioning

confidence: 99%

Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity

2016

View full text Add to dashboard Cite

Abstract:The discovery and generation of biocatalysts with extended catalytic versatilities are of immense relevance in both chemistry and biotechnology. An enhanced atomistic understanding of enzyme promiscuity, a mechanism through which living systems acquire novel catalytic functions and specificities by evolution, would thus be of central interest. Using esterase-catalyzed amide bond hydrolysis as a model system, we pursued a simplistic in silico discovery program aiming for the identification of enzymes with an internal backbone hydrogen bond acceptor that could act as a reaction specificity shifter in hydrolytic enzymes. Focusing on stabilization of the rate limiting transition state of nitrogen inversion, our mechanism-guided approach predicted that the acyl hydrolase patatin of the α/β phospholipase fold would display reaction promiscuity. Experimental analysis confirmed previously unknown high amidase over esterase activity displayed by the first described esterase machinery with a protein backbone hydrogen bond acceptor to the reacting NH-group of amides. The present work highlights the importance of a fundamental understanding of enzymatic reactions and its potential for predicting enzyme scaffolds displaying alternative chemistries amenable to further evolution by enzyme engineering.

show abstract

“…73 The proposed mechanism for synthesis of TMM is as follows in Scheme 4. 33, 67, 72 Pks13 has two ACP domains.…”

Section: Trehalose Utilization Pathways (Tups)mentioning

confidence: 99%

Targeting the trehalose utilization pathways ofMycobacterium tuberculosis

Thanna

Sucheck

2016

Med. Chem. Commun.

View full text Add to dashboard Cite

Tuberculosis (TB) is an epidemic disease and the growing burden of multidrug-resistant (MDR) TB world wide underlines the need to discover new drugs to treat the disease. Mycobacterium tuberculosis (Mtb) is the etiological agent of most cases of TB. Mtb is difficult to treat, in part, due to the presence of a sturdy hydrophobic barrier that prevents penetration of drugs through the cell wall. Mtb can also survive in a non-replicative state for long periods of time avoiding the action of common antibiotics. Trehalose is an essential metabolite in mycobacteria since it plays key roles in cell wall synthesis, transport of cell wall glycolipids, and energy storage. It is also known for its stress protective roles such as: protection from desiccation, freezing, starvation and osmotic stress in bacteria. In this review we discuss the drug discovery efforts against enzymes involved in the trehalose utilization pathways (TUPs) and their likelihood of becoming drug targets.

show abstract

Biochemical and Structural Study of the Atypical Acyltransferase Domain from the Mycobacterial Polyketide Synthase Pks13

Cited by 44 publications

References 57 publications

Architecture of the polyketide synthase module: surprises from electron cryo-microscopy

Architecture of the polyketide synthase module: surprises from electron cryo-microscopy

Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity

Targeting the trehalose utilization pathways ofMycobacterium tuberculosis

Contact Info

Product

Resources

About