PPIase activities of AquaCyp293 and AquaCyp300.

<p>(A) Kinetics of <i>cis</i>/<i>trans</i> isomerization of 3 μM Abz-Ala-Ala-Pro-Phe-pNa followed by fluorescence at 416 nm, without enzyme (black), with 8 nM (blue), 12 nM (green), 16 nM (dark blue) and 20 nM (red) AquaCyp293. (B) Refolding kinetics of RCM-T1 in the presence of increasing concentrations of AquaCyp293, 0 nM (black), 10 nM (blue), 300 nM (green) and 750 nM (red). The kinetics of refolding of 0.1 μM RCM-T1 in 0.1 M Tris/HCl pH 8.0; 2 M NaCl were measured at 15°C in the presence of various concentrations of AquaCyp293. (C, D) Catalytic efficiencies of AquaCyp293 (○) and AquaCyp300 (□) for (C) the <i>cis</i>/<i>trans</i> isomerization of Abz-Ala-Ala-Pro-Phe-<i>p</i>NA and (D) the refolding of RCM-T1. The measured proline-limited refolding rate constants <i>k</i>_app are shown as a function of the PPIase concentration. The <i>k</i>_cat/<i>K</i>_m values derived from the slopes are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157070#pone.0157070.t001" target="_blank">Table 1</a>.</p

Dimerization of AquaCyp300.

<p>(A) Surface representation of the dimeric AquaCyp300 crystal structure, one protomer is colored in light the other in dark grey, the N-terminal-, insertion, and C-terminal structural elements in one protomer are colored blue, green and red, respectively, active site residues of both protomers are colored yellow. (B) Sequence conservation [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157070#pone.0157070.ref083" target="_blank">83</a>] within the AquaCyp300 family is mapped onto a cartoon representation of AquaCyp300 protomer2. Residues that are highly conserved (e.g. Phe258, Phe259, Phe260) are shown in blue, sequences with lower identity are shown in white and red.</p

Domain structure and conservation of AquaCyp.

<p>The cyclophilin domains are shown in grey, the additonal N-terminal-, insertion, and C-terminal structural elements are colored in blue, green and red, respectively. The indicated amino acids are conserved among cyclophilins. The two invariant cysteine residues in AquaCyp are shown in bold. Extension and insertion regions are indicated above the sequences.</p

Active site structure of AquaCyp293 (A) and AquaCyp300 (B) in comparison to hCyp18 (C) and EcCypB (D).

<p>Residues that contribute to the active site of the cyclophilin family are labeled and shown in stick representation. (E) Conservation of PPIase active site residues. Residue numbering according to hCyp18 (C).</p

Statistics on diffraction data and structure refinement of the AquaCyp293 and AquaCyp300.

<p>Statistics on diffraction data and structure refinement of the AquaCyp293 and AquaCyp300.</p

Crystal structures of AquaCyp293 and AquaCyp300.

<p>Domain architecture and topology of AquaCyp293 (A) and AquaCyp300 (B). The cyclophilin fold in grey consists of an eight stranded-antiparallel β-barrel and two α-helices covering the top and the bottom of the barrel. The disulfide bridge (yellow) is shown in stick representation. The additional N-terminal-, insertion, and C-terminal structural elements are coloroured in blue, green and red, respectively.</p

Catalytic efficiencies of AquaCyp293 and AquaCyp300 for prolyl isomerization in peptide and protein substrates.

<p>Catalytic efficiencies of AquaCyp293 and AquaCyp300 for prolyl isomerization in peptide and protein substrates.</p