Structural and functional characterization of a putative carbonic anhydrase from Geobacillus kaustophilus reveals its cambialistic function

Sridharan, Upasana; Ragunathan, Preethi; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Kumarevel, Thirumananseri; Ponnuraj, Karthe

doi:10.1016/j.bbrc.2021.02.036

Cited by 4 publications

(9 citation statements)

References 23 publications

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“…The quaternary structure of the purified protein was investigated by SEC indicating that in our experimental conditions, BpsγCA is trimeric ( Fig. 2 A), as reported for γ-CAs previously characterized [55] , [67] , [68] , [69] , [70] , [71] , [72] , [73] , [74] . Circular dichroism experiments carried out at 25 °C allowed us to estimate a preponderant content of β-sheet secondary structure (29 %).…”

Section: Resultssupporting

confidence: 65%

“…Interaction between two adjacent monomers in the trimer is very extensive, with a buried surface at the monomer–monomer interface of about 1917 Å 2 . Similarly to the previously characterized γ-CA family members [55] , [67] , [68] , [69] , [70] , [71] , [72] , [73] , [74] , there are three active sites per trimer located at the monomer–monomer interfaces ( Fig. 3 B) in large clefts characterized by a highly hydrophobic base and polar edges ( Fig.…”

Section: Resultsmentioning

confidence: 62%

“…The superposition of BpsγCA with the γ-CAs previously characterized (see Table 2 ), namely RicA [55] , Zn-Cap [72] , and the γ-CAs from Methanosarcina thermophila (Cam) [67] , E. c oli (Yrda) [71] , Thermosynechococcus elongatus (TeCcmM) [70] , the Discovery Deep Brine Pool (CA_D) [69] , Thermus thermophilus HB8 (TtCA) [68] , and Geobacillus k austophilus (Cag) [74] , revealed a substantial conservation of their three-dimensional structure, with the highest similarity detected with Zn-Cap (identity = 43.5 %, r.m.s.d. = 0.6 Å) [72] and RicA (identity = 48.9 %, r.m.s.d.…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei

Fiore

Luca²,

Langella³

et al. 2022

Computational and Structural Biotechnology Journal

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Section: Resultssupporting

confidence: 65%

Section: Resultsmentioning

confidence: 62%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei

Fiore

Luca²,

Langella³

et al. 2022

Computational and Structural Biotechnology Journal

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“…Carbonic anhydrase proteins that catalyze hydration of carbon dioxide are involved in a wide range of fundamental biological processes in plants, fungi, and bacteria ( Supuran, 2016 ; Dostál et al, 2018 ). However, the structural evolution has resulted in classifying these enzymes into distinct classes ( Elleuche and Pöggeler, 2009 ; Supuran, 2018a ; Dreyer et al, 2020 ; Sridharan et al, 2021 ) and the conservation of each class in different domains of life makes them a potential target for controlling biotic diseases ( Nishimori et al, 2007 ; Innocenti et al, 2009 ; Syrjänen et al, 2015 ; Supuran, 2016 ; Urbański et al, 2020 ). Despite being well-studied in humans, higher eukaryotes, β-CAs have been analyzed in only few fungal species ( Kim et al, 2020 ; Urbański et al, 2020 ; Vullo et al, 2020 ; Supuran and Capasso, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

“…For this assay, Tris buffer and BSA were used as a blank and negative control, respectively, carbonic anhydrase from bovine erythrocytes (BCA; Merck, Shanghai, China) was used as a positive control. The improve method from Wilbur and Anderson was used ( Sridharan et al, 2021 ) that involved monitoring of time taken during the pH change of 12 mM Tris Buffer at 0°C from 8.3 to 6.3 in the presence of carbonic anhydrase. CO 2 bubbled double distilled water was used as the substrate for this reaction.…”

Section: Methodsmentioning

confidence: 99%

Contribution of the Mitochondrial Carbonic Anhydrase (MoCA1) to Conidiogenesis and Pathogenesis in Magnaporthe oryzae

Dang

Batool

et al. 2022

Front. Microbiol.

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The interconversion of CO2 and HCO3− catalyzed by carbonic anhydrases (CAs) is a fundamental biochemical process in organisms. During mammalian–pathogen interaction, both host and pathogen CAs play vital roles in resistance and pathogenesis; during planta–pathogen interaction, however, plant CAs function in host resistance but whether pathogen CAs are involved in pathogenesis is unknown. Here, we biologically characterized the Magnaporthe oryzae CA (MoCA1). Through detecting the DsRED-tagged proteins, we observed the fusion MoCA1 in the mitochondria of M. oryzae. Together with the measurement of CA activity, we confirmed that MoCA1 is a mitochondrial zinc-binding CA. MoCA1 expression, upregulated with H2O2 or NaHCO3 treatment, also showed a drastic upregulation during conidiogenesis and pathogenesis. When MoCA1 was deleted, the mutant ΔMoCA1 was defective in conidiophore development and pathogenicity. 3,3′-Diaminobenzidine (DAB) staining indicated that more H2O2 accumulated in ΔMoCA1; accordingly, ATPase genes were downregulated and ATP content decreased in ΔMoCA1. Summarily, our data proved the involvement of the mitochondrial MoCA1 in conidiogenesis and pathogenesis in the rice blast fungus. Considering the previously reported HCO3− transporter MoAE4, we propose that MoCA1 in cooperation with MoAE4 constitutes a HCO3− homeostasis-mediated disease pathway, in which MoCA1 and MoAE4 can be a drug target for disease control.

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Rational engineering of a highly active and resilient α‐carbonic anhydrase from the hydrothermal vent species Persephonella hydrogeniphila

Manyumwa,

Zhang,

Jers

et al. 2024

The FEBS Journal

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Carbonic anhydrases (CAs) are ideal catalysts for carbon dioxide sequestration in efforts to alleviate climate change. Here, we report the characterisation of three α‐CAs that originate from the thermophilic bacteria Persephonella hydrogeniphila (PhyCA), Persephonella atlantica (PaCA), and Persephonella sp. KM09‐Lau‐8 (PlauCA) isolated from hydrothermal vents. The three α‐Cas, showing high sequence similarities, were produced in Escherichia coli, purified and characterised. Surprisingly, they revealed very different behaviours with regards to their thermostability profiles. PhyCA presented a more stable thermostability profile amongst the three, thus we chose it for rational engineering to improve it further. PhyCA's residue K88, a proton transfer residue in α‐CAs, was mutated to His, Ala, Gln and Tyr. A 4‐fold activity improvement was noted for variants K88H and K88Q at 30 °C, owing to the higher proton transfer efficiency of the replacement proton transfer residues. K88Q also proved more stable than PhyCA. K88Y did not increase activity, but notably increased thermal stability, with this enzyme variant retaining 50% of its initial activity after incubation for 1 h at 90 °C. Removal of the two main proton shuttles (variant H85A_K88A) resulted in diminished activity of the enzyme. Molecular dynamics simulations performed for PhyCA and all its variants revealed differences in residue fluctuations, with K88A resulting in a general reduction in root mean square fluctuation (RMSF) of active site residues as well as most of the CA's residues. Its specific activity and stability in turn increased compared to the wild type.

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Structural and functional characterization of a putative carbonic anhydrase from Geobacillus kaustophilus reveals its cambialistic function

Cited by 4 publications

References 23 publications

Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei

Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei

Contribution of the Mitochondrial Carbonic Anhydrase (MoCA1) to Conidiogenesis and Pathogenesis in Magnaporthe oryzae

Rational engineering of a highly active and resilient α‐carbonic anhydrase from the hydrothermal vent species Persephonella hydrogeniphila

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