Membrane Anchoring and Intervesicle Transfer of a Derivative of the Antibiotic Moenomycin A

Anikin, Alexej; Buchynskyy, Andrij; Kempin, Uwe; Stembera, Katka; Welzel, Peter; Lantzsch, Gabriela

doi:10.1002/(sici)1521-3773(19991216)38:24<3703::aid-anie3703>3.0.co;2-1

Cited by 29 publications

(24 citation statements)

References 7 publications

(24 reference statements)

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“…After lyophilization, pure 2 (16 mg, 64 %) was obtained. 1 H NMR (partly broad signals by micelle formation, [27] Expression and preparation of PBP 1b: The protein was purified from the membrane fraction of E. coli strain JM 109 (K-12 recA1 D(lac-proAB) endA1 gyrA96 thi1 hsdR17 supE44 relA1 F' [tra-D36, proAB , lacI q , lacZDM15]) carrying the plasmid pJP13. This strain expresses the structural gene of PBP 1b of E. coli.…”

Section: Discussionmentioning

confidence: 99%

“…In the purification process proteins were analyzed by SDS-PAGE and visualized by silver staining and Western blotting with anti-PBP 1b antibodies. For the competition SPR experiments, [26] moenomycin derivative 6 (Scheme 3) [27] was immobilized on a sensor chip surface, which carried N-hydroxysuccinimideactivated carboxylic acid groups.…”

Section: Spr Competition Experimentsmentioning

confidence: 99%

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A Surface Plasmon Resonance Analysis of the Interaction between the Antibiotic Moenomycin A and Penicillin‐Binding Protein 1b

et al. 2002

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The antibiotic moenomycin A inhibits the biosynthesis of peptidoglycan, the main structural polymer of the bacterial cell wall. The inhibition is based on a reversible binding of the antibiotic to one of the substrate binding sites in enzymes such as penicillin-binding protein (PBP) 1b. A novel assay based on surface plasmon resonance (SPR) has been established that can be used to investigate selective binding of the moenomycin sugar moiety and other transglycosylase inhibitors to this enzyme. Suitable ligands were prepared from moenomycin A and coupled to SPR sensor surfaces. Moenomycin analogues with structural variations were used to perform competitive SPR experiments with PBP 1b. The SPR results confirm for the first time that the trisaccharide fragment of moenomycin A (C-E-F-G-H-I) is the minimal structure that possesses all moieties sufficient for biological activity and for affinity towards PBP 1b. The method seems to be appropriate for use in screens for transglycosylase inhibitors that bind to the moenomycin-binding site of the enzyme.

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Section: Discussionmentioning

confidence: 99%

Section: Spr Competition Experimentsmentioning

confidence: 99%

A Surface Plasmon Resonance Analysis of the Interaction between the Antibiotic Moenomycin A and Penicillin‐Binding Protein 1b

et al. 2002

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“…The recent description of x-ray crystal structures of the PBP2 extracellular domain from S. aureus (18) and the TG domain of PBP1a from Aquifex aeolicus (19) have provided invaluable structural insights into the plausible mechanism of cell wall peptidoglycan polymerization. Lacking, however, is information regarding the role of the TM domain of PBPs in catalysis, although it has been speculated to interact with the lipid moiety of moenomycin or lipid II (20,21). To address this question, protein constructs containing different domains of PBP1b from Escherichia coli were expressed and purified with suitable detergents [see supporting information (SI) Materials and Methods and SI Fig.…”

Section: Domain Requirement Of Moenomycin Binding To Class a Pbps (Bimentioning

confidence: 99%

Domain requirement of moenomycin binding to bifunctional transglycosylases and development of high-throughput discovery of antibiotics

Cheng

Sung

Liao

et al. 2008

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

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Moenomycin inhibits bacterial growth by blocking the transglycosylase activity of class A penicillin-binding proteins (PBPs), which are key enzymes in bacterial cell wall synthesis. We compared the binding affinities of moenomycin A with various truncated PBPs by using surface plasmon resonance analysis and found that the transmembrane domain is important for moenomycin binding. Full-length class A PBPs from 16 bacterial species were produced, and their binding activities showed a correlation with the antimicrobial activity of moenomycin against Enterococcus faecalis and Staphylococcus aureus. On the basis of these findings, a fluorescence anisotropy-based high-throughput assay was developed and used successfully for identification of transglycosylase inhibitors.penicillin-binding proteins ͉ transglycosylase inhibitors ͉ fluorescence anisotropy M any common bacterial pathogens, such as Staphylococcus aureus, Streptococcus pneumoniae, and Enterococcus faecalis, have become multidrug-resistant and have emerged as a public health concern, creating an urgent need for new antibiotics.The bacterial cell wall, or its peptidoglycan synthesis pathway, has been targeted for the development of antibacterial agents (1). The synthesis of peptidoglycan consists of several steps, including the formation of lipid I and lipid II, followed by the final transglycosylation and transpeptidation of lipid II to form peptidoglycan (1, 2). Many current antibiotics are ␤-lactam derivatives that target transpeptidation. To our knowledge, no medicines have yet been developed to inhibit the transglycosylation process. The only known potent inhibitors for transglycosylase (TG) are moenomycin complexes (flavomycin), including moenomycin A (Moe A) (Fig. 1A, compound 1), A12, C1, C3, and C4 (3, 4). Among these, Moe A is the most abundant agent in its family (3, 4). The unique antibacterial properties of Moe A have prompted chemists to synthesize moenomycin fragments and derivatives (5-7) in an attempt to develop new antibiotics. Recently, the total synthesis of Moe A (8), and of its biosynthesis pathway (9), has been reported. However, due to poor bioavailability, flavomycin is currently used only as a growth promoter in animal feeds (10).The characterization of class A penicillin-binding proteins (PBPs) and the identification of TG inhibitors require functional PBP and lipid II as the substrate for the enzyme. However, the limited availability of lipid II has hampered the development of effective enzymatic assays for identification of inhibitors. As a result, the majority of the screening methods that are used to search for TG inhibitors, including the low-throughput methods that use surface plasmon resonance (SPR) (12) or radioactive assays (13-16), rely mainly on moenomycin (11). Development of a TG activity assay that is amenable to high-throughput screening (HTS) is thus desirable for inhibitor identification.In this study, we compared the binding of moenomycin to various truncated PBPs and concluded that the transmembrane (TM) domain is c...

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“…From structureactivity relationship analysis, it has been found that moenomycin trisaccharides containing the units C, E, F, G, H, and I are the smallest fragments that show antibiotic activity in vivo, whereas moenomycin disaccharides containing the units E-I still act as transglycosylase inhibitors in vitro [3, 7-9, 12, 25]. The isoprenoidlike C 25 -alcohol [26], named moenocinol, seems to be required for the binding of the moenomycin antibiotics to the cytoplasmic membrane [27], whereas the units E, F, and H are concluded to form a polar binding epitope that is recognized by the donor binding site of the transglycosylases [3, 7-9, 19, 28]. In addition to the five known constituents of the Flavomycin complex, structurally related moenomycin-type antibiotics, including the prasinomycins, the diumycins (macarbomycins), ensanchomycin, prenomycin, teichomycin, pholipomycin, AC326-␣, 11837 RP, 8036 RP (quebemecin), and 19402 RP, have been found [3,23,29,30].…”

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Characterization of moenomycin antibiotic complex by multistage MALDI-IT/RTOF-MS and ESI-IT-MS

Zehl

Pittenauer

Rizzi

et al. 2006

J. Am. Soc. Mass Spectrom.

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Flavomycin is a commercially available antimicrobial growth promoter and an authorized additive for feeding stuffs in the EU and in the USA. As most antibiotically active products biosynthesized by microorganisms, it contains not only a single active compound but is a complex mixture of structurally closely related substances. Multistage matrix-assisted laser desorption/ionization-ion trap/reflectron time-of-flight mass spectrometry (MALDI-IT/ RTOF-MS) and liquid chromatography-electrospray ionization-ion trap-mass spectrometry (LC-ESI-IT-MS) were utilized for a detailed analysis of the constituents of the Flavomycin complex based on low-energy collision induced dissociation (CID). An optimal sample preparation for negative ion vacuum MALDI-MS for this compound class was developed. The MALDI-IT/RTOF-MS 2 and -MS 3 analysis starting with the precursor [M Ϫ H] Ϫ ions of these interesting phosphoglycolipids, named moenomycins, yielded a large variety of product ions that facilitated the structural characterization of this class of compounds. Based on the derived CID fragmentation pathway of the five known major constituents, namely moenomycin A, moenomycin A 12 , moenomycin C 4 , moenomycin C 3 . and moenomycin C 1 , four not yet described moenomycin-type constituents could be characterized. They were assigned as 4F-demethyl-6E-O-de-␤-D-glucopyranosyl-moenomycin A, 6B-N-de(2-hydroxy-5-oxo-1-cyclopenten-1-yl)-moenomycin A, 6B-hydroxy-6B-de[N-(2-hydroxy-5-oxo-1-cyclopenten-1-yl)amino]-moenomycin A, and 6C-hydroxy-moenomycin A. In addition, a moenomycin A carrying an oxygen in the moenocinol-group was found, which is most probably a chemical degradation product. These new compounds were verified by LC-ESI-IT-MS. (J Am Soc Mass

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Membrane Anchoring and Intervesicle Transfer of a Derivative of the Antibiotic Moenomycin A

Cited by 29 publications

References 7 publications

A Surface Plasmon Resonance Analysis of the Interaction between the Antibiotic Moenomycin A and Penicillin‐Binding Protein 1b

A Surface Plasmon Resonance Analysis of the Interaction between the Antibiotic Moenomycin A and Penicillin‐Binding Protein 1b

Domain requirement of moenomycin binding to bifunctional transglycosylases and development of high-throughput discovery of antibiotics

Characterization of moenomycin antibiotic complex by multistage MALDI-IT/RTOF-MS and ESI-IT-MS

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