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DiB-splits: nature-guided design of a novel fluorescent labeling split system.

ABSTRACT: Fluorogen-activating proteins (FAPs) are innovative fluorescent probes combining advantages of genetically-encoded proteins such as green fluorescent protein and externally added fluorogens that allow for highly tunable and on demand fluorescent signaling. Previously, a panel of green- and red-emitting FAPs has been created from bacterial lipocalin Blc (named DiBs). Here we present a rational design as well as functional and structural characterization of the first self-assembling FAP split system, DiB-splits. This new system decreases the size of the FAP label to ~8-12?kDa while preserving DiBs' unique properties: strong increase in fluorescence intensity of the chromophore upon binding, binding affinities to the chromophore in nanomolar to low micromolar range, and high photostability of the protein-ligand complex. These properties allow for use of DiB-splits for wide-field, confocal, and super-resolution fluorescence microscopy. DiB-splits also represent an attractive starting point for further design of a protein-protein interaction detection system as well as novel FAP-based sensors.

SUBMITTER: Bozhanova NG

PROVIDER: S-EPMC7338535 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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DiB-splits: nature-guided design of a novel fluorescent labeling split system.

Bozhanova Nina G NG Gavrikov Alexey S AS Mishin Alexander S AS Meiler Jens J

Scientific reports 20200706 1

Fluorogen-activating proteins (FAPs) are innovative fluorescent probes combining advantages of genetically-encoded proteins such as green fluorescent protein and externally added fluorogens that allow for highly tunable and on demand fluorescent signaling. Previously, a panel of green- and red-emitting FAPs has been created from bacterial lipocalin Blc (named DiBs). Here we present a rational design as well as functional and structural characterization of the first self-assembling FAP split syst ...[more]

PMID: 32632329

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Project description:ObjectivesTo evaluate the feasibility of a newly proposed minimally invasive split-thickness flap design without vertical-releasing incisions for vertical bone regeneration performed in either a simultaneous or staged approach and to analyze the prevalence of adverse events during postoperative healing.Materials and methodsFollowing preparation of a split-thickness flap and bilaminar elevation of the mucosa and underlying periosteum, the alveolar bone was exposed over the defects, vertical GBR was performed by means of a titanium-reinforced high-density polytetrafluoroethylene membrane combined with particulated autogenous bone (AP) and bovine-derived xenograft (BDX) in 1:1 ratio. At 9 months after reconstructive surgery, vertical and horizontal hard tissue gain was evaluated based on clinical and radiographic examination.ResultsTwenty-four vertical alveolar ridge defects in 19 patients were treated with vertical GBR. In case of 6 surgical sites, implant placement was performed at the time of the GBR (simultaneous group); in the remaining 18 surgical, sites implant placement was performed 9 months after the ridge augmentation (staged group). After uneventful healing in 23 cases, hard tissue fill was detected in each site. Direct clinical measurements confirmed vertical and horizontal hard tissue gain averaging 3.2 ± 1.9 mm and 6.5 ± 0.5 mm respectively, in the simultaneous group and 4.5 ± 2.2 mm and 8.7 ± 2.3 mm respectively, in the staged group. Additional radiographic evaluation based on CBCT data sets in the staged group revealed mean vertical and horizontal hard tissue fill of 4.2 ± 2.0 mm and 8.5 ± 2.4 mm. Radiographic volume gain was 1.1 ± 0.4 cm3.ConclusionVertical GBR consisting of a split-thickness flap and using titanium-reinforced non-resorbable membrane in conjunction with a 1:1 mixture of AP+BDX may lead to a predictable vertical and horizontal hard tissue reconstruction.Clinical relevanceThe used split-thickness flap design may represent a valuable approach to increase the success rate of vertical GBR, resulting in predicable hard tissue regeneration, and favorable wound healing with low rate of membrane exposure.

Dataset Information

DiB-splits: nature-guided design of a novel fluorescent labeling split system.

Publications

DiB-splits: nature-guided design of a novel fluorescent labeling split system.

Similar Datasets

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