A fusion of the Bacteroides fragilis ferrous iron import proteins reveals a role for FeoA in stabilizing GTP-bound FeoB

Sestok, Alex E.; Brown, Janae B.; Juliet, O.; O’Sullivan, Sean M.; Garcin, Elsa D.; Deredge, Daniel; Smith, Aaron T.

doi:10.1016/j.jbc.2022.101808

Cited by 10 publications

(7 citation statements)

References 75 publications

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“…Sequence coverage determination and HDX‐MS experiments were performed as described 58 , 59 with the following conditions. FVIII and CR.7–8 samples were buffer exchanged into 20 mM imidazole pH 7.3, 10 mM CaCl 2 , and 500 mM NaCl.…”

Section: Methodsmentioning

confidence: 99%

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Characterization of interaction between blood coagulation factor VIII and LRP1 suggests dynamic binding by alternating complex contacts

Chun

Kurasawa

Olivares

et al. 2022

Journal of Thrombosis and Haemostasis

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Background Deficiency in blood coagulation factor VIII (FVIII) results in life‐threating bleeding (hemophilia A) treated by infusions of FVIII concentrates. To improve disease treatment, FVIII has been modified to increase its plasma half‐life, which requires understanding mechanisms of FVIII catabolism. An important catabolic actor is hepatic low density lipoprotein receptor‐related protein 1 (LRP1), which also regulates many other clinically significant processes. Previous studies showed complexity of FVIII site for binding LRP1. Objectives To characterize binding sites between FVIII and LRP1 and suggest a model of the interaction. Methods A series of recombinant ligand‐binding complement‐type repeat (CR) fragments of LRP1 including mutated variants was generated in a baculovirus system and tested for FVIII interaction using surface plasmon resonance, tissue culture model, hydrogen–deuterium exchange mass spectrometry, and in silico. Results Multiple CR doublets within LRP1 clusters II and IV were identified as alternative FVIII‐binding sites. These interactions follow the canonical binding mode providing major binding energy, and additional weak interactions are contributed by adjacent CR domains. A representative CR doublet was shown to have multiple contact sites on FVIII. Conclusions FVIII and LRP1 interact via formation of multiple complex contacts involving both canonical and non‐canonical binding combinations. We propose that FVIII‐LRP1 interaction occurs via switching such alternative binding combinations in a dynamic mode, and that this mechanism is relevant to other ligand interactions of the low‐density lipoprotein receptor family members including LRP1.

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Section: Methodsmentioning

confidence: 99%

“…Sequence coverage determination and HDX-MS experiments were performed as described 58,59 with the following conditions.…”

Section: Hydrogen Deuterium Exchange-mass Spectrometrymentioning

confidence: 99%

Characterization of interaction between blood coagulation factor VIII and LRP1 suggests dynamic binding by alternating complex contacts

Chun

Kurasawa

Olivares

et al. 2022

Journal of Thrombosis and Haemostasis

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“…pylori, but in E. coli and Bacteroides fragilis, it acts as a regulatory accessory to FeoB, the transmembrane transport protein. , In B. fragilis, FeoA is proposed to bind to FeoB to activate Fe 2+ transport and to dissociate and reduce Fe 2+ transport when needs are satisfied.…”

Section: Resultsmentioning

confidence: 99%

“…The putative Fe 2+ regulatory peptide FeoA has not been characterized from H. pylori, but in E. coli and Bacteroides fragilis, it acts as a regulatory accessory to FeoB, the transmembrane transport protein. 27,28 In B. fragilis, FeoA is proposed to bind to FeoB to activate Fe 2+ transport and to dissociate and reduce Fe 2+ transport when needs are satisfied. The C39Y mutation in H. pylori FeoA can be postulated to assist in Fe 2+ transport to satisfy the induced radical SAM enzymes of menaquinone synthesis.…”

Section: Acs Infectiousmentioning

confidence: 99%

MAT Gain of Activity Mutation in Helicobacter pylori Is Associated with Resistance to MTAN Transition State Analogues

et al. 2023

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Helicobacter pylori is found in the gut lining of more than half of the world’s population, causes gastric ulcers, and contributes to stomach cancers. Menaquinone synthesis in H. pylori relies on the rare futalosine pathway, where H. pylori 5′-methylthioadenosine nucleosidase (MTAN) is proposed to play an essential role. Transition state analogues of MTAN, including BuT-DADMe-ImmA (BTDIA) and MeT-DADMe-ImmA (MTDIA), exhibit bacteriostatic action against numerous diverse clinical isolates of H. pylori with minimum inhibitory concentrations (MIC’s) of <2 ng/mL. Three H. pylori BTDIA-resistant clones were selected under increasing BTDIA pressure. Whole genome sequencing showed no mutations in MTAN. Instead, resistant clones had mutations in metK, methionine adenosyltransferase (MAT), feoA, a regulator of the iron transport system, and flhF, a flagellar synthesis regulator. The mutation in metK causes expression of a MAT with increased catalytic activity, leading to elevated cellular S-adenosylmethionine. Metabolite analysis and the mutations associated with resistance suggest multiple inputs associated with BTDIA resistance. Human gut microbiome exposed to MTDIA revealed no growth inhibition under aerobic or anaerobic conditions. Transition state analogues of H. pylori MTAN have potential as agents for treating H. pylori infection without disruption of the human gut microbiome or inducing resistance in the MTAN target.

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“…Although research on the bacterial Feo system has primarily focused on Gammaproteobacteria species, alternative architectures of Feo have been identified in other groups ( Sestok et al, 2018 ). For example, the commensal species Bacteroides fragilis has a single Feo protein containing a fusion of FeoA and FeoB homolog domains ( Sestok et al, 2022 ). A common feature found when comparing the Feo system among species is that FeoA and FeoB orthologs (or their corresponding domains) are nearly universal, while FeoC is poorly conserved, being present in about 5% of bacterial proteomes, predominantly within the Gamma group ( Gómez-Garzón et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Divide and conquer: genetics, mechanism, and evolution of the ferrous iron transporter Feo in Helicobacter pylori

Gómez-Garzón

Payne

2023

Front. Microbiol.

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IntroductionFeo is the most widespread and conserved system for ferrous iron uptake in bacteria, and it is important for virulence in several gastrointestinal pathogens. However, its mechanism remains poorly understood. Hitherto, most studies regarding the Feo system were focused on Gammaproteobacterial models, which possess three feo genes (feoA, B, and C) clustered in an operon. We found that the human pathogen Helicobacter pylori possesses a unique arrangement of the feo genes, in which only feoA and feoB are present and encoded in distant loci. In this study, we examined the functional significance of this arrangement.MethodsRequirement and regulation of the individual H. pylori feo genes were assessed through in vivo assays and gene expression profiling. The evolutionary history of feo was inferred via phylogenetic reconstruction, and AlphaFold was used for predicting the FeoA-FeoB interaction.Results and DiscussionBoth feoA and feoB are required for Feo function, and feoB is likely subjected to tight regulation in response to iron and nickel by Fur and NikR, respectively. Also, we established that feoA is encoded in an operon that emerged in the common ancestor of most, but not all, helicobacters, and this resulted in feoA transcription being controlled by two independent promoters. The H. pylori Feo system offers a new model to understand ferrous iron transport in bacterial pathogens.

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A fusion of the Bacteroides fragilis ferrous iron import proteins reveals a role for FeoA in stabilizing GTP-bound FeoB

Cited by 10 publications

References 75 publications

Characterization of interaction between blood coagulation factor VIII and LRP1 suggests dynamic binding by alternating complex contacts

Characterization of interaction between blood coagulation factor VIII and LRP1 suggests dynamic binding by alternating complex contacts

MAT Gain of Activity Mutation in Helicobacter pylori Is Associated with Resistance to MTAN Transition State Analogues

Divide and conquer: genetics, mechanism, and evolution of the ferrous iron transporter Feo in Helicobacter pylori

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