The crystal structure of Escherichia coli MoeA, a protein from the molybdopterin synthesis pathway

Schrag, Joseph D.; Huang, Weijun; Sivaraman, J.; Smith, Christopher J.; Plamondon, J.; Larocque, Robert; Matte, Allan; Cygler, Mirosław

doi:10.1006/jmbi.2001.4771

Cited by 50 publications

(38 citation statements)

References 58 publications

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“…proteins. Crystal structures and biochemical experiments have indicated that MoeA exists as a dimer (10,11) and MogA as a trimer (6), whereas MobA is monomeric (4, 5). MobB exists as a dimer according to gel filtration experiments (22).…”

Section: Resultsmentioning

confidence: 99%

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In VivoInteractions between Gene Products Involved in the Final Stages of Molybdenum Cofactor Biosynthesis inEscherichia coli

Magalon

Frixon

Pommier

et al. 2002

Journal of Biological Chemistry

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The final stages of bacterial molybdenum cofactor (Moco) biosynthesis correspond to molybdenum chelation and nucleotide attachment onto an unique and ubiquitous structure, the molybdopterin. Using a bacterial two-hybrid approach, here we report on the in vivo interactions between MogA, MoeA, MobA, and MobB implicated in several distinct although linked steps in Escherichia coli. Numerous interactions among these proteins have been identified. Somewhat surprisingly, MobB, a GTPase with a yet unclear function, interacts with MogA, MoeA, and MobA. Probing the effects of various mo. mutations on the interaction map allowed us (i) to distinguish Moco-sensitive interactants from insensitive ones involving MobB and (ii) to demonstrate that molybdopterin is a key molecule triggering or facilitating MogA-MoeA and MoeA-MobA interactions. These results suggest that, in vivo, molybdenum cofactor biosynthesis occurs on protein complexes rather than by the separate action of molybdenum cofactor biosynthetic proteins.

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Section: Resultsmentioning

confidence: 99%

“…However, in vitro mixing of purified MoeA and MogA proteins under differ- In Vivo Interactions among mo. Gene Productsent conditions did not form detectable complexes, leading to the assumption that either these proteins do not interact or that a factor was missing under these conditions (10). Hence, the effect of mo.…”

Section: Discussionmentioning

confidence: 99%

In VivoInteractions between Gene Products Involved in the Final Stages of Molybdenum Cofactor Biosynthesis inEscherichia coli

Magalon

Frixon

Pommier

et al. 2002

Journal of Biological Chemistry

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“…The crystal structure of MoeA shows it to exist as a homodimer, and it is likely that proteins with sequence similarity to MoeA do so as well (Schrag et al, 2001;Xiang et al, 2001). We have observed that the LIN-46 protein strongly interacts with itself in a yeast two hybrid assay, further suggesting LIN-46 forms multimers (L. Tang and E.G.M., unpublished).…”

Section: Lin-46 May Form a Scaffold For A Multiprotein Assemblymentioning

confidence: 87%

TheC. elegansheterochronic genelin-46affects developmental timing at two larval stages and encodes a relative of the scaffolding protein gephyrin

et al. 2004

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lin-46 is required prior to the third stage for normal adult cell fates, suggesting that it acts once to control fates at both stages, and that it affects adult fates through the let-7 branch of the heterochronic pathway. Interestingly, lin-46 encodes a protein homologous to MoeA of bacteria and the C-terminal domain of mammalian gephyrin, a multifunctional scaffolding protein. Our findings suggest that the LIN-46 protein acts as a scaffold for a multiprotein assembly that controls developmental timing, and expand the known roles of gephyrin-related proteins to development.

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“…Each monomer is composed of four distinct domains that in the following discussion will be referred to as the sub-domains of Geph-E. The E domain of the homologous plant protein Cnx1 (28) also dimerizes, and Geph-E has been postulated to be dimeric as well (29,30). Based on the distinct oligomerization potentials of the G and E domains, the formation of a hexagonal gephyrin scaffold located underneath the postsynaptic membrane was suggested (29), which would allow efficient clustering of large numbers of GlyRs (1).…”

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Biochemical Characterization of the High Affinity Binding between the Glycine Receptor and Gephyrin

Schrader

Kim

Winking

et al. 2004

Journal of Biological Chemistry

107

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Gephyrin is an essential and instructive molecule for the formation of inhibitory synapses. Gephyrin binds directly to the large cytoplasmic loop located between transmembrane helices three and four of the ␤-subunit of the glycine receptor and to microtubules, thus promoting glycine receptor (GlyR) anchoring to the cytoskeleton and clustering in the postsynaptic membrane. Besides its structural role, gephyrin is involved in the biosynthesis of the molybdenum cofactor that is essential for all molybdenum-dependent enzymes in mammals. Gephyrin can be divided into an N-terminal trimeric G domain and a C-terminal E domain, which are connected by a central linker region. Here we have studied the in vitro interaction of gephyrin and its domains with the large cytoplasmic loop of the GlyR ␤-subunit (GlyR␤-loop). Binding of gephyrin to the GlyR is exclusively mediated by the E domain, and the binding site was mapped to one of its sub-domains (residues 496 -654). By using isothermal titration calorimetry, a high affinity (K d ‫؍‬ 0.2-0.4 M) and low affinity (K d ‫؍‬ 11-30 M) binding site for the GlyR␤-loop was found on holo-gephyrin and the E domain, respectively, with a binding stoichiometry of two GlyR␤-loops per E domain in both cases. Binding of the GlyR␤-loop does not change the oligomeric state of either full-length gephyrin or the isolated E domain.Efficient transmission of synaptic signals in the central nervous system is dependent on high local concentrations of neurotransmitter ion channels in the postsynaptic membrane of excitatory and inhibitory synapses. Glycine is an important inhibitory neurotransmitter, and its postsynaptic receptors are an essential component of inhibitory nerve terminals (1-3). Glycine receptors (GlyRs) 1 are pentameric anion channels generally consisting of two different subunits with an ␣ 3 ␤ 2 composition (4). Crucial for proper functioning of GlyRs is their clustering that generates a high packing density of neuroreceptors in the postsynaptic membrane. Gephyrin, a highly expressed neuronal protein, was found to co-localize with glycine (5, 6) and GABA A receptors (7,8). Gephyrin is crucial for the clustering of both types of inhibitory neuroreceptors (9) and anchors these receptors to the subsynaptic cytoskeleton. A direct binding of gephyrin to microtubules was shown (10), which is possibly mediated by a "tau" motif (11). So far no direct interaction with microfilaments has been demonstrated, but pharmacological studies indicate a function of actin filaments in determining the cluster size (12). Therefore, it is not surprising that binding of gephyrin to key regulators of microfilament dynamics such as profilin I, neuronal profilin IIa, and microfilament adaptors of the Mena/VASP family including neuronal Mena has been reported recently (13). The binding site of gephyrin on GlyR has been mapped to a stretch of 18 amino acids located within the large cytoplasmic loop connecting transmembrane helices three and four of the GlyR ␤-subunit (GlyR␤-loop) (14), but the binding site on ge...

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The crystal structure of Escherichia coli MoeA, a protein from the molybdopterin synthesis pathway

Cited by 50 publications

References 58 publications

In VivoInteractions between Gene Products Involved in the Final Stages of Molybdenum Cofactor Biosynthesis inEscherichia coli

In VivoInteractions between Gene Products Involved in the Final Stages of Molybdenum Cofactor Biosynthesis inEscherichia coli

TheC. elegansheterochronic genelin-46affects developmental timing at two larval stages and encodes a relative of the scaffolding protein gephyrin

Biochemical Characterization of the High Affinity Binding between the Glycine Receptor and Gephyrin

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