The DNA sequence of the sulfate activation locus from Escherichia coli K-12.

Leyh, Thomas S.; Vogt, Thomas; Suo, Ya

doi:10.1016/s0021-9258(19)50034-5

Cited by 87 publications

(17 citation statements)

References 48 publications

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“…The native enzyme is a tetramer of heterodimers (13); each dimer is composed of a GTPase, CysN (53 kDa), and its target, CysD (35 kDa) (14). Activated sulfate is an essential intermediate in the biosynthesis of reduced sulfur metabolites in aerobic bacteria (15).…”

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confidence: 99%

Conformational Change Rate-Limits GTP Hydrolysis: The Mechanism of the ATP Sulfurylase−GTPase

Jiang

Leyh

1998

Biochemistry

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The fluorescent GTP analogues 3'-O-(N-methylanthraniloyl)-2'-deoxyguanosine 5'-(beta, gamma-imidotriphosphate) (mGMPPNP) and 3'-O-(N-methylanthraniloyl)-2'-deoxy-GTP (mGTP) were used to demonstrate that an enzyme isomerization precedes and rate-limits beta,gamma-bond cleavage in the catalytic cycle of the ATP sulfurylase-GTPase, from E. coli K-12. The binding of mGMPPNP to the E.AMP.PPi complex of ATP sulfurylase is biphasic, indicating that an isomerization occurs in the binding reaction. The isomerization mechanism was assigned based on the results of the enzyme concentration dependence of the observed rate constants, kobs, for both phases of the binding reaction, and sequential-mixing, nucleotide release experiments. The isomerization occurs after, and is driven by, the addition of mGMPPNP. Values were determined for each of the rate constants associated with the two-step kinetic model used in the interpretation of the results. A comparison of the enzyme concentration dependence of kobs for the hydrolysis and binding reactions reveals that the rate constants for the corresponding steps of these two reactions are extremely similar. The virtually identical rate constants for isomerization and beta, gamma-bond scission strongly suggest that isomerization rate-limits bond breaking. The implications of these finding for GTPase/target interactions and the mechanism of energetic linkage in the ATP sulfurylase system are discussed.

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confidence: 99%

Conformational Change Rate-Limits GTP Hydrolysis: The Mechanism of the ATP Sulfurylase−GTPase

Jiang

Leyh

1998

Biochemistry

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“…The expression of enzymes ATPS and APSK is fundamentally required for biocatalytic synthesis of PAPS from ATP (Figure a). The bacterial ATPS forms a tetramer composed of two heterodimers and requires guanosine 5′-triphosphate for activation, while the fungal ATPS is directly expressed as an active form . Therefore, we expressed the codon-optimized ATPS gene from the yeast K.…”

Section: Resultsmentioning

confidence: 99%

Closed-Loop System Driven by ADP Phosphorylation from Pyrophosphate Affords Equimolar Transformation of ATP to 3′-Phosphoadenosine-5′-phosphosulfate

Wang

Huang

et al. 2021

ACS Catal.

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3′-Phosphoadenosine-5′-phosphosulfate (PAPS) is a universal sulfate group donor for all biological sulfation reactions in living organisms. Ambitions to biomanufacture sulfate-containing compounds such as heparin and chondroitin sulfate also promote the study on PAPS in vitro synthesis. However, the established enzymatic synthesis of PAPS faces hurdles of the natural low theoretical transformation rate of 50% (two ATP to one PAPS) and high cost. Here, we developed a PAPS synthesis route with 100% theoretical transformation rate which affords equimolar transformation of ATP to PAPS. By fusing the identified adenosine 5′-triphosphate sulfurylase and adenosine 5′-phosphosulfate kinase from different species, we created an artificial active bifunctional enzyme to directly convert ATP to PAPS. To maximize the conversion from ATP to PAPS, a polyphosphate (polyP)-dependent ATP regeneration system was designed and engineered by screening polyP kinases which consumes the low-cost polyP as the phosphate donor. In addition, we found that PPi could be used as the phosphate donor for phosphorylating ADP to ATP by polyP kinases. After demonstration of the wide distribution of PPi kinase activity in polyP kinases, a closed-loop ATP regeneration route was thereupon created to convert one ATP to one PAPS in theory. Using PPi as the phosphate donor, the conversion rate of ATP to PAPS reached 92.3%. The efficient enzymatic route that is constructed here for PAPS synthesis with low cost would boost the biosynthesis of sulfated compounds and peptides.

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“…Bacteria have a large repertoire of selenocysteine-containing proteins, and both SelD and CysD play vital roles in their biosynthesis. While the role of selenoproteins in cellular stress management is well documented (53), CysD is also involved in the first step of sulfite formation during the assimilatory reduction of sulfate to sulfide (54). Since ROS is a major driver of cell death during thermal stress (55, 56), the highly reduced sulfide species produced during sulfate reduction can potentially defend microbial cells amid high heat (57).…”

Section: Discussionmentioning

confidence: 99%

Thermal endurance by a hot-spring-dwelling phylogenetic relative of the mesophilic Paracoccus

Mondal

Roy

Chatterjee

et al. 2022

Preprint

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High temperature growth/survival was revealed in a phylogenetic relative (strain SMMA_5) of the mesophilic Paracoccus isolated from the 78-85°C water of a Trans- Himalayan sulfur-borax spring. After 12 h at 50°C, or 45 minutes at 70°C, in mineral salts thiosulfate (MST) medium, SMMA_5 retained ∼2% colony-forming units (CFUs), whereas comparator Paracoccus had 1.5% and 0% CFU left at 50°C and 70°C respectively. After 12 h at 50°C, the thermally-conditioned sibling SMMA_5_TC exhibited ∼1.5 time increase in CFU-count; after 45 minutes at 70°C, SMMA_5_TC had 7% of the initial CFU-count intact. 1000-times diluted Reasoner’s 2A medium, and MST supplemented with lithium, boron or glycine-betaine (solutes typical of the SMMA_5 habitat), supported higher CFU-retention/CFU-growth than MST. With or without lithium/boron/glycine-betaine in MST, a higher percentage of cells always remained viable (cytometry data), compared with what percentage remained capable of forming single colonies (CFU data). SMMA_5, compared with other Paracoccus, contained 335 unique genes, mostly for DNA replication/recombination/repair, transcription, secondary metabolites biosynthesis/transport/catabolism, and inorganic ion transport/metabolism. It’s also exclusively enriched in cell wall/membrane/envelope biogenesis, and amino acid metabolism, genes. SMMA_5 and SMMA_5_TC mutually possessed 43 nucleotide polymorphisms, of which 18 were in protein-coding genes with 13 nonsynonymous and seven radical amino acid replacements. Such biochemical and biophysical mechanisms could be involved in thermal stress mitigation which streamline the cells’ energy and resources towards system-maintenance and macromolecule-stabilization, thereby relinquishing cell-division for cell-viability. Thermal conditioning apparently helped memorize those potential metabolic states which are crucial for cell-system maintenance, while environmental solutes augmented the indigenous stability-conferring mechanisms.IMPORTANCEFor a holistic understanding of microbial life’s high-temperature adaptation it is imperative to explore the biology of the phylogenetic relatives of mesophilic bacteria which get stochastically introduced to geographically and geologically diverse hot spring systems by local geodynamic forces. Here, in vitro endurance of high heat up to the extent of growth under special (habitat-inspired) conditions was discovered in a hot- spring-dwelling phylogenetic relative of the mesophilic Paracoccus species. Thermal conditioning, extreme oligotrophy, metabolic deceleration, presence of certain habitat- specific inorganic/organic solutes, and typical genomic specializations were found to be the major enablers of this conditional (acquired) thermophilicity. Feasibility of such phenomena across the taxonomic spectrum can well be paradigm-changing for the established scopes of microbial adaptation to the physicochemical extremes. Applications of conditional thermophilicity in microbial process biotechnology may be far reaching and multi-faceted.

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The DNA sequence of the sulfate activation locus from Escherichia coli K-12.

Cited by 87 publications

References 48 publications

Conformational Change Rate-Limits GTP Hydrolysis: The Mechanism of the ATP Sulfurylase−GTPase

Conformational Change Rate-Limits GTP Hydrolysis: The Mechanism of the ATP Sulfurylase−GTPase

Closed-Loop System Driven by ADP Phosphorylation from Pyrophosphate Affords Equimolar Transformation of ATP to 3′-Phosphoadenosine-5′-phosphosulfate

Thermal endurance by a hot-spring-dwelling phylogenetic relative of the mesophilic Paracoccus

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