Structural insights into catalytic and substrate binding mechanisms of the strategic EndA nuclease from Streptococcus pneumoniae

Moon, A.F.; Midon, Marika; Meiß, Gregor; Pingoud, Alfred; London, Robert E.; Pedersen, Lars C.

doi:10.1093/nar/gkq1152

Cited by 36 publications

(77 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the authors' opinion [84], the catalytic mecha nism of phosphodiester bond hydrolysis by endonu clease EndA is similar to that proposed for endonu cleases Sm [70,85] and NucA [82] (Fig. 18).…”

Section: Nonspecific Endonucleasessupporting

confidence: 69%

“…The Q186A mutation is less deleterious to the catalysis than the E205A mutation. In the authors' opinion [84], this is due to the fact that Gln186 interacts with only one water molecule from the inner coordination sphere of the divalent metal Fig. 18.…”

Section: Nonspecific Endonucleasesmentioning

confidence: 95%

“…The crystal structure of ββα metal nuclease EndA from S. pneumoniae was determined by X ray diffraction at 1.75 Å resolution (Fig. 10) [84]. A comparison of the EndA structure with the structures of other nucleases belonging to the same family revealed a conserved core containing a β sheet and a ββα metal fragment that form the catalytic site.…”

Section: Nonspecific Endonucleasesmentioning

confidence: 99%

See 2 more Smart Citations

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

et al. 2012

View full text Add to dashboard Cite

Endonucleases (EC 3.1) are enzymes of the hydrolase class that catalyze the hydrolytic cleavage of deoxyribonucleic and ribonucleic acids at any region of the polynucleotide chain. Endonucleases are widely used both in biotechnological processes and in veterinary medicine as antiviral agents. Medical appli cations of endonucleases in human cancer therapy hold promise. The results of X ray diffraction studies of the spatial organization of endonucleases and their complexes and the mechanism of their action are ana lyzed and generalized. An analysis of the structural studies of this class of enzymes showed that the specific binding of enzymes to nucleic acids is characterized by interactions with nitrogen bases and the nucleotide backbone, whereas the nonspecific binding of enzymes is generally characterized by interactions only with the nucleic acid backbone. It should be taken into account that the specificity can be modulated by metal ions and certain low molecular weight organic compounds. To test the hypotheses about specific and non specific nucleic acid binding proteins, it is necessary to perform additional studies of atomic resolution three dimensional structures of enzyme-nucleic acid complexes by methods of structural biology.

show abstract

Section: Nonspecific Endonucleasessupporting

confidence: 69%

Section: Nonspecific Endonucleasesmentioning

confidence: 95%

Section: Nonspecific Endonucleasesmentioning

confidence: 99%

See 1 more Smart Citation

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

et al. 2012

View full text Add to dashboard Cite

show abstract

“…For the DNase activity assays shown in Fig. 3B, purified wild-type CPS-6 (0.06 -2 M) was incubated with 60 nM 48-mer ssDNA (5Ј-ACGCTGCCGAATTCTGGCGTTAGGAGATA-CCGATAAGCTTCGGCTTAA) or dsDNA (above 48-mer ssDNA annealed with its complementary sequence), both of which were 32 P-labeled at the 5Ј-end in the optimal CPS-6 buffer. For the DNase activity assays shown in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…For the DNase activity assays shown in Fig. 3C, purified wild-type protein (0.25-1 M) was incubated with either 10 nM 11-mer ssDNA (5Ј-AACCTTACAAC-3Ј) or ssRNA (5Ј-AAC-CUUACAAC-3Ј), both of which were labeled at the 5Ј-end with 32 P, in the optimal CPS-6 buffer. For the DNase activity assays shown in Fig.…”

Section: Methodsmentioning

confidence: 99%

Structural Insights into Apoptotic DNA Degradation by CED-3 Protease Suppressor-6 (CPS-6) from Caenorhabditis elegans

Lin

Nakagawa

Lin

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: CPS-6 (EndoG) degrades chromosomal DNA during apoptosis. Results: The crystal structure of C. elegans CPS-6 was determined, and the DNA binding and cleavage mechanisms by CPS-6 were revealed. Conclusion: The DNase activity of CPS-6 is positively correlated with its pro-cell death activity. Significance: This study improves our general understanding of DNA hydrolysis by ␤␤␣-metal finger nucleases and the process of apoptotic DNA fragmentation.

show abstract

Chemical Rescue of Active Site Mutants of S. pneumoniae Surface Endonuclease EndA and Other Nucleases of the HNH Family by Imidazole

Midon¹,

Gimadutdinow²,

Meiß³

et al. 2012

ChemBioChem

Self Cite

View full text Add to dashboard Cite

The His-Asn-His (HNH) motif characterizes the active sites of a large number of different nucleases such as homing endonucleases, restriction endonucleases, structure-specific nucleases and, in particular, nonspecific nucleases. Several biochemical studies have revealed an essential catalytic function for the first amino acid of this motif in HNH nucleases. This histidine residue was identified as the general base that activates a water molecule for a nucleophilic attack on the sugar phosphate backbone of nucleic acids. Replacement of histidine by an amino acid such as glycine or alanine, which lack the catalytically active imidazole side chain, leads to decreases of several orders of magnitude in the nucleolytic activities of members of this nuclease family. We were able, however, to restore the activity of HNH nuclease variants (i.e., EndA (Streptococcus pneumoniae), SmaNuc (Serratia marcescens) and NucA (Anabaena sp.)) that had been inactivated by His→Gly or His→Ala substitution by adding excess imidazole to the inactive enzymes in vitro. Imidazole clearly replaces the missing histidine side chain and thereby restores nucleolytic activity. Significantly, this chemical rescue could also be observed in vivo (Escherichia coli). The in vivo assay might be a promising starting point for the development of a high-throughput screening system for functional EndA inhibitors because, unlike the wild-type enzyme, the H160G and H160A variants of EndA can easily be produced in E. coli. A simple viability assay would allow inhibitors of EndA to be identified because these would counteract the toxicities of the chemically rescued EndA variants. Such inhibitors could be used to block the nucleolytic activity of EndA, which as a surface-exposed enzyme in its natural host destroys the DNA scaffolds of neutrophil extracellular traps (NETs) and thereby allows S. pneumoniae to escape the innate immune response.

show abstract

Structural insights into catalytic and substrate binding mechanisms of the strategic EndA nuclease from Streptococcus pneumoniae

Cited by 36 publications

References 48 publications

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

Structural aspects of catalytic mechanisms of endonucleases and their binding to nucleic acids

Structural Insights into Apoptotic DNA Degradation by CED-3 Protease Suppressor-6 (CPS-6) from Caenorhabditis elegans

Chemical Rescue of Active Site Mutants of S. pneumoniae Surface Endonuclease EndA and Other Nucleases of the HNH Family by Imidazole

Contact Info

Product

Resources

About