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110 results for “promiscuous”
Fig. 7 in Site-directed mutagenesis of β sesquiphellandrene synthase enhances enzyme promiscuity
Fig. 7. Mass spectra and retention index of the hydroxylated sesquiterpene generated by PmSTS mutants L454G and L454A. NIST library mass spectra of (A) sesquicineole, (B) cis-sesquisabinene hydrate, (C) trans-sesquisabinene hydrate, and (D) α-bisabolol are shown at bottom of each respective sample mass spectra.
Fig. 4 in Site-directed mutagenesis of β sesquiphellandrene synthase enhances enzyme promiscuity
Fig. 4. Structural analysis by circular dichroism (CD) of PmSTSΔ24WT and L454 mutants. Far-UV CD spectra (190–260 nm) of PmSTSΔ24 WT, PmSTSΔ24Y390S/L454G, PmSTSΔ24L454G and PmSTSΔ24L454A are shown. Buffer used was 20 mM Tris pH 8.0, 100 mM NaCl 2 mM βME. Protein concentration was 10 μM. Further secondary structure variations in percentages for each L454 mutant and WT were determined using the JASCO SSE program are presented in Table S2.
Fig. 3 in Site-directed mutagenesis of β sesquiphellandrene synthase enhances enzyme promiscuity
Fig. 3. Analysis of monomeric fraction of PmSTS Wild Type and mutant proteins using (A) 12% SDS PAGE and (B) 10% Native PAGE. The SDS PAGE showed high purity of PmSTS protein were obtained for wild type and for all mutants except for PmSTSΔ24W286A and PmSTSΔ24Y390S. The native PAGE showed several species of PmSTSΔ24WT, PmSTSΔ24V466E, and PmSTSΔ24Y390S/L454G that can be resolved on a 10% native gel. However, only one species of PmSTSΔ24L454G and PmSTSΔ24L454A was resolved on a 10% native gel. Smearing bands were observed for PmSTSΔ24W286A and PmSTSΔ24Y390S on the 10% native gel, probably due to the low purity as judged by 12% SDS PAGE.
Fig. 1 in Site-directed mutagenesis of β sesquiphellandrene synthase enhances enzyme promiscuity
Fig. 1. Superimposition of PmSTS homology model (green) with the crystal structure of Nicotiana tabacum 5-epi-aristolochene synthase (NtEAS, PDB ID:3M01). NtEAS has a FPP ligand analog (2-trans, 6-trans)-2- fluorofarnesyl diphosphate (FPF) bound in the active site (pink) shows an RMSD of 1.4 Å over 518 aligned Cα atoms. The Mg2+ ions in the active site of NtEAS are shown in yellow spheres. PmSTS and NtEAS share ~38% sequence identify. The putative active residues of PmSTS (W286, Y390, L454 and V466) shown heavier sticks were selected for mutagenesis studies. The structure superimposition was carried out using SSM in COOT (Emsley et al., 2010). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Fig. 2 in Site-directed mutagenesis of β sesquiphellandrene synthase enhances enzyme promiscuity
Fig. 2. Size exclusion chromatography (SEC) profiles of affinity chromatography purified wild-type and mutant proteins. The chromatography was performed on a HiLoad Superdex column 200 PG. Detection was at 280 nm. (A) PmSTSΔ24 WT, (B) PmSTSΔ24 V466E, (C) PmSTSΔ24 W286A, (D) PmSTSΔ24 Y390S, (E) PmSTSΔ24L454A, (F) PmSTSΔ24L454G, and (G) PmSTSΔ24Y390S/L454G were run over a size exclusion column to verify the oligomeric status of generated proteins. Elution profiles of β-sesquiphellandrene synthase proteins are represented together with protein standard: bovine thyroglobulin (670 kDa), bovine gamma globulin (158 kDa), ovalbumin (44 kDa), myoglobin (17 kDa), vitamin B12 (1.35 kDa). The peaks eluted at ~48 mL correspond to protein aggregated during the purification process, and peaks eluted at ~80 mL correspond to the monomeric size of PmSTSΔ24 protein (65 kDa).
Data from: Innovative females are more promiscuous in great tits (Parus major)
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Data from: The promiscuous and the chaste: frequent allopolyploid speciation and its genomic consequences in American daisies (Melampodium sect. Melampodium; Asteraceae)
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Data from: Strategic promiscuity helps avoid inbreeding at multiple levels in a cooperative breeder where both sexes are philopatric
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Data from: Sex-specific estimates of dispersal show female philopatry and male dispersal in a promiscuous amphibian, the alpine salamander (Salamandra atra)
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Data from: Heritability of plumage colour morph variation in a wild population of promiscuous, long-lived Australian magpies
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Data from: Promiscuity resolves constraints on social mate choice imposed by population viscosity
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Data from: Social structure of the harem-forming promiscuous fruit bat, Cynopterus sphinx, is the harem truly important?
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Data from: Promiscuous mating in the harem-roosting fruit bat, Cynopterus sphinx
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Data from: Widely naturalized species are not more promiscuous in the use of different nitrogen forms, but benefit more from inorganic nitrogen
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Maintaining genetic integrity with high promiscuity: Frequent hybridization with low introgression in multiple hybrid zones of Melocactus(Cactaceae)
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Data sets for compound promiscuity analysis and predictions
<p>Deposited are three data sets containing compounds with multi- or single-target activity, which were assembled from the PubChem BioAssay database for promiscuity predictions [1]. The design and composition of these data sets are described in the original publication [1] and a forthcoming data note detailing the deposition. A brief summary of the data structure is provided in the readme.txt file accompanying the data sets.</p>
Data from: Active site loop conformation regulates promiscuous activity in a lactonase from Geobacillus kaustophilus HTA426
Enzyme promiscuity is a prerequisite for fast divergent evolution of biocatalysts. A phosphotriesterase-like lactonase (PLL) from Geobacillus kaustophilus HTA426 (GkaP) exhibits main lactonase and promiscuous phosphotriesterase activities. To understand its catalytic and evolutionary mechanisms, we investigated a "hot spot" in the active site by saturation mutagenesis as well as X-ray crystallographic analyses. We found that position 99 in the active site was involved in substrate discrimination. One mutant, Y99L, exhibited 11-fold improvement over wild-type in reactivity (kcat/Km) toward the phosphotriesterase substrate ethyl-paraoxon, but showed 15-fold decrease toward the lactonase substrate δ-decanolactone, resulting in a 157-fold inversion of the substrate specificity. Structural analysis of Y99L revealed that the mutation causes a ~6.6 Å outward shift of adjacent loop 7, which may cause increased flexibility of the active site and facilitate accommodation and/or catalysis of organophosphate substrate. This study provides for the PLL family an example of how the evolutionary route from promiscuity to specificity can derive from very few mutations, which promotes alteration in the conformational adjustment of the active site loops, in turn draws the capacity of substrate binding and activity.
Data from: Exposure to parasites increases promiscuity in a freshwater snail
Under the Red Queen hypothesis, outcrossing can produce genetically variable progeny, which may be more resistant, on average, to locally adapted parasites. Mating with multiple partners may enhance this resistance by further increasing the genetic variation among offspring. We exposed Potamopyrgus antipodarum to the eggs of a sterilising, trematode parasite and tested whether this altered mating behaviour. We found that exposure to parasites increased the number of snail mating pairs and the total number of different mating partners for both males and females. Thus our results suggest that, in host populations under parasite-mediated selection, exposure to infective propagules increases the rate of mating and the number of mates.
Understanding the enzyme (S)-Norcoclaurine Synthase promiscuity to aldehydes and ketones
<p>Files of the work 'Understanding the enzyme (S)-Norcoclaurine Synthase promiscuity to aldehydes and ketones' where was studied the application of methods in silico to understand the reactivity of compounds</p>
Figure 3 in Who's your daddy? Paternity testing reveals promiscuity and multiple paternity in the carnivorous marsupial Dasyurus maculatus (Marsupialia: Dasyuridae)
Figure 3. Relatedness within litters with one (open bars) and two sires (solid bars).
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Annotated Behaviour and Observability Dataset (ABODe)
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