Recombinant FimH Adhesin Demonstrates How the Allosteric Catch Bond Mechanism Can Support Fast and Strong Bacterial Attachment in the Absence of Shear

Thomas, Wendy E.; Carlucci, Laura A.; Yakovenko, Olga; Interlandi, Gianluca; Trong, I. Le; Aprikian, Pavel; Magala, Pearl; Larson, Lydia; Sledneva, Yulia; Tchesnokova, Veronika; Stenkamp, Ronald E.; Sokurenko, Evgeni V.

doi:10.1016/j.jmb.2022.167681

Cited by 4 publications

(30 citation statements)

References 86 publications

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“…This is consistent with experimental data showing that in the absence of mannose both FimH and FimH FocH are likely to be found in LAS with LD and PD docked to each other. 10 Despite no differences in overall flexibility, previous studies by the author on protein complexes have provided evidence that mutations near a binding interface can often induce changes in the network of inter-protein (in this case, inter-domain) contacts. 23,24 For this reason, I analysed the persistence of inter-domain contacts between LD and PD, and compared FimH to…”

Section: Comparison Of Backbone Flexibility and Inter-domain Contactsmentioning

confidence: 99%

“…Evidence for such an equilibrium shift in the absence of shear has been provided by a recent study by us where wild-type FimH was compared to variants that are more easily activated. 10 In particular, in one of the variants LD of FimH was fused in a recombinant manner to a structurally altered PD where a segment of 16 residues was replaced with a related sequence from FocH, a naturally occurring homologue of FimH with different lectin specificity. Overall, this fusion results in 10 mutations in PD (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

“…Overall, this fusion results in 10 mutations in PD (Figure 1). I refer here to the combination of LD from FimH and PD with the aminoacid sequence replacement from FocH as FimH FocH (in a previous study, 10 it has been termed LD-PD FocH ). The X-ray structure of FimH FocH crystallized in the absence of mannose reveals that LD and PD are bound to each other with LD in LAS in a very similar fashion as in wild-type FimH 10 (Figure 1a,b).…”

Section: Introductionmentioning

confidence: 99%

“…However, in functional assays FimH FocH was found in the presence of mannose to be activated and displayed HAS-like phenotype even in the absence of shear while wild-type FimH required shear-induced tensile force for sustained activation. 10 The observation that FimH FocH is activated under static conditions while wild-type FimH requires shear despite their crystallographic structures having no significant differences suggests a differential activation mechanism between FimH FocH and FimH. This motivates studying the dynamics of the activation mechanism at atomic level of detail.…”

Section: Introductionmentioning

confidence: 99%

“…A previous study by us found that the mutation A188D in FimH FocH , which is known to be activating, 11,12 weakens the LD-PD interface by disrupting hydrophobic inter-domain side-chain interactions. 10 However, the mutations in FimH FocH may also affect inter-domain electrostatic interactions, which are thought to be stronger than hydrophobic contacts. Hence, to thoroughly understand how the FimH FocH mutations alter the LD-PD interface it is necessary to investigate, besides hydrophobic-driven side chain contacts, also electrostatic interactions including intra-PD as the mutations may alter networks of hydrogen bonds and salt bridges.…”

Section: Introductionmentioning

confidence: 99%

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Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Interlandi

2023

Preprint

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The bacterial adhesin FimH is a model for the study of protein allostery because its structure has been resolved in multiple configurations, including the active and the inactive state. FimH consists of a pilin domain (PD) that anchors it to the rest of the fimbria and an allosterically regulated lectin domain (LD) that binds mannose on the surface of infected cells. Under normal conditions, the two domains are docked to each other and LD binds mannose weakly. However, in the presence of tensile force generated by shear the domains separate and conformational changes propagate across LD resulting in a stronger bond to mannose. Recently, the crystallographic structure of a variant of FimH has been resolved, called FimH , where PD contains 10 mutations near the inter-domain interface. Although the X-ray structures of FimH and FimH are almost identical, experimental evidence shows that FimH is activated even in the absence of shear. Here, molecular dynamics simulations combined with the Jarzinski equality were used to investigate the discrepancy between the crystallographic structures and the functional assays. The results indicate that the free energy barrier of the unbinding process between LD and PD is drastically reduced in FimH . Rupture of an inter-domain hydrogen bond involving R166 constitutes a rate limiting step of the domains separation process and occurs more readily in FimH than FimH. In conclusion, the mutations in FimH shift the equilibrium towards an equal occupancy of bound and unbound states for LD and PD by reducing a rate limiting step.

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Section: Comparison Of Backbone Flexibility and Inter-domain Contactsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Interlandi

2023

Preprint

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Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Interlandi

2023

Proteins

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The bacterial adhesin FimH is a model for the study of protein allostery because its structure has been resolved in multiple configurations, including the active and the inactive state. FimH consists of a pilin domain (PD) that anchors it to the rest of the fimbria and an allosterically regulated lectin domain (LD) that binds mannose on the surface of infected cells. Under normal conditions, the two domains are docked to each other and LD binds mannose weakly. However, in the presence of tensile force generated by shear the domains separate and conformational changes propagate across LD resulting in a stronger bond to mannose. Recently, the crystallographic structure of a variant of FimH has been resolved, called , where PD contains 10 mutations near the inter‐domain interface. Although the X‐ray structures of FimH and are almost identical, experimental evidence shows that is activated even in the absence of shear. Here, molecular dynamics simulations combined with the Jarzynski equality were used to investigate the discrepancy between the crystallographic structures and the functional assays. The results indicate that the free energy barrier of the unbinding process between LD and PD is drastically reduced in . Rupture of inter‐domain hydrogen bonds involving R166 constitutes a rate limiting step of the domain separation process and occurs more readily in than FimH. In conclusion, the mutations in shift the equilibrium toward an equal occupancy of bound and unbound states for LD and PD by reducing a rate limiting step.

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Multiscale Allostery: Basic Mechanisms and Versatility in Diagnostics and Drug Design

Berezovsky

Nussinov²

2022

Journal of Molecular Biology

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Recombinant FimH Adhesin Demonstrates How the Allosteric Catch Bond Mechanism Can Support Fast and Strong Bacterial Attachment in the Absence of Shear

Cited by 4 publications

References 86 publications

Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Rate limiting step of the allosteric activation of the bacterial adhesin FimH investigated by molecular dynamics simulations

Multiscale Allostery: Basic Mechanisms and Versatility in Diagnostics and Drug Design

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