4 research outputs found
Genetic structures located upstream of <i>parF</i> and <i>parG.</i>
<p>A. The direct repeats within the pMET1 putative <i>parH</i>-like locus are shown in red. The diagram also shows the −35 and −10 sequences, as well as the inverted repeats (arrows). The inverted repeat within the putative <i>parH</i> locus is shown in blue. The beginning of the ParF amino acid sequence including the deviant Walker motif A and motif A' are shown. B. Logo plot <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Crooks1" target="_blank">[60]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Schneider1" target="_blank">[61]</a> of a multiple alignment of the direct repeats shown in red.</p
Genetic map of pMET1 and comparison to plasmid pCRY and chromosomal elements.
<p>A. The genetic maps of pMET1 and pCRY are compared showing the homologous regions. The arrows indicate genes locations and orientation. Genes with different functions are shown with different colors and if the genes in the different structures shown are homologus they are represented with the same colors. Yellow: mobilization; green: replication and partition; red: antibiotic resistance; purple: virB/pilX-like; blue: transposition; grey: unknown. Since pCRY is smaller than pMET1, to represent it in circular form a dotted line was added to fill the gap. Solid line represents non-homologous DNA. B. Comparison of a pMET1 region with chromosomal HPIs or ICEs is shown using a linearized version of the plasmid. The HPIs shown are those from <i>E. coli</i> ECOR31 (HPI<sub>ECOR31</sub>) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Schubert1" target="_blank">[43]</a>, <i>K. pneumoniae</i> NTUH-K2044 (ICE<sub>Kp1</sub>) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Lin1" target="_blank">[44]</a>, and <i>Y. pestis</i> KIM (HPI<sub>Yp</sub>)<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Schubert1" target="_blank">[43]</a>. The diagram shows the HP core regions, which are not at scale and are represented as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001800#pone.0001800-Schubert1" target="_blank">[43]</a>, and the RB-HPIs. The sequence described in this manuscript has been deposited in GenBank, accession number is EU383016.</p
Replication region of pMET1.
<p>A. The bar shows a genetic map of the pMET1 replication region and the GC content plot generated using a window size of 100 bp on top. Recombinant clones were obtained by inserting the indicated fragments into pCR2.1 or ligated to the pUC4K <i>aph</i> cassette. The ability to be maintained in <i>E. coli</i> C2110 (a <i>polA</i> mutant) of the recombinant plasmids made using pCR2.1 as vector is indicated to the right by a + or − sign. The ability to generate kanamycin resistant colonies in <i>E. coli</i> TOP10 of the indicated fragments when ligated to the <i>aph</i> cassette from pUC4K is also represented by a + or − sign. B. BLASTP comparison of the amino acid sequences of the putative RepA proteins from pMET1 and pCRY.</p
Discovery of IWP-051, a Novel Orally Bioavailable sGC Stimulator with Once-Daily Dosing Potential in Humans
In
recent years, soluble guanylate cyclase (sGC, EC 4.6.1.2) has
emerged as an attractive therapeutic target for treating cardiovascular
diseases and diseases associated with fibrosis and end-organ failure.
Herein, we describe our design and synthesis of a series of 4-hydroxypyrimidine
sGC stimulators starting with an internally discovered lead. Our efforts
have led to the discovery of IWP-051, a molecule that achieves good
alignment of potency, stability, selectivity, and pharmacodynamic
effects while maintaining favorable pharmacokinetic properties with
once-daily dosing potential in humans