Of 50637630 A. A least-squares superposition of subunits with LSQKAB [41] gives an

Of 50637630 A. A least-squares superposition of subunits with LSQKAB [41] gives an r.m.s.d. (root-mean-square ?deviation) of 0.57 A for 90 Ca atoms, which shows there are no major conformational differences between the two subunits. It is noteworthy that such a low value was 4 IBP biological activity obtained in the absence of NCS restraints. The total surface area of a subunit, calculated with PISA [38], is ??approximately 7400 A2 of which 1700 A2 are buried within the dimer. Therefore, about 23 of the surface area of each monomer is involved in dimerization. The free energy of dissociation (DGdiss) is estimated as 19.4 kcal mol21, and suggests that this assembly is thermodynamically stable, consistent with the observation of a stable dimer in solution. Similar values are observed for other SCAN structures. For example, the interface area and DGdiss for the Znf24 dimer (PDB code 3LHR) are 23 and 21.8 kcal mol21, respectively. At present there 16574785 are four SCAN domain structures in the PDB, two crystal structures and two determined by solution NMR. BIBS39 cost sequence conservation of these four with human PEG3-SCAN is presented in Fig. 2. The superposition of the PEG3-SCAN dimer onto these other dimers reveals an overall structural conservation (Fig. 4), with calculated r.m.s.d. values presented in Table 2. The largest deviations among SCAN structures occur at the N- and Cterminal ends, which show higher flexibility than the core, and a4, which is positioned away from the dimer interface. The r.m.s.d.Figure 3. Overall structure of PEG3-SCAN. The homodimer is shown as ribbons with one subunit green, the partner purple. The Nand C- termini as well as the five a-helices of each monomer are labeled accordingly. doi:10.1371/journal.pone.0069538.gvalues for alignments with the SCAN domain dimers of Znf42 and ?Znf174 show higher variation, more than 1.0 A greater, than for the X-ray structures, because of the greater uncertainties associated with the NMR structures and that the fit involves an average of 20 conformers that represent their NMR derived structures.Residues Forming the SCAN Dimer InterfaceThe human PEG3-SCAN homodimer is held together by an extensive network of hydrogen-bonding, salt-bridge interactions and van der Waals forces. Even though the overall sequence identity among the five known SCAN structures is only 40?0 (Fig. 2), the key residues located at the dimer interface and that contribute to inter-subunit associations are conserved. TheSCAN Domain of PEGTable 2. Structure and sequence similarity between PEG3-SCAN and other SCAN domains.?R.m.s.d (A) 1.57 1.51 2.85 2.Protein name Zfp206 Znf24 Znf42 ZnfPDB codes 4E6S 3LHR 2FI2 1Y7QR.m.s.d alignment length 157 164 155Sequence identity ( ) 38 48 35These included crystal structures of Zfp206 and Znf24, and solution NMR structures of Znf42 and Znf174. R.m.s.d. calculations were carried out with PDBeFold using secondary structure matching [49] with the PEG3-SCAN dimer in the superposition. Sequence alignment was performed with ClustalW2 using residues 40?30 of the full-length PEG3 against the core of the SCAN domain, as well as 2? flanking residues, of other proteins. doi:10.1371/journal.pone.0069538.tmajority of these intermolecular contacts are formed between a1 and a2 (the N-terminal sub-domain) of one subunit and a3 on the C-terminal sub-domain of the partner. Helices a2 and a3 show the highest amino acid conservation when comparing the sequences of these known SCAN domain structures and the conserved residues cont.Of 50637630 A. A least-squares superposition of subunits with LSQKAB [41] gives an r.m.s.d. (root-mean-square ?deviation) of 0.57 A for 90 Ca atoms, which shows there are no major conformational differences between the two subunits. It is noteworthy that such a low value was obtained in the absence of NCS restraints. The total surface area of a subunit, calculated with PISA [38], is ??approximately 7400 A2 of which 1700 A2 are buried within the dimer. Therefore, about 23 of the surface area of each monomer is involved in dimerization. The free energy of dissociation (DGdiss) is estimated as 19.4 kcal mol21, and suggests that this assembly is thermodynamically stable, consistent with the observation of a stable dimer in solution. Similar values are observed for other SCAN structures. For example, the interface area and DGdiss for the Znf24 dimer (PDB code 3LHR) are 23 and 21.8 kcal mol21, respectively. At present there 16574785 are four SCAN domain structures in the PDB, two crystal structures and two determined by solution NMR. Sequence conservation of these four with human PEG3-SCAN is presented in Fig. 2. The superposition of the PEG3-SCAN dimer onto these other dimers reveals an overall structural conservation (Fig. 4), with calculated r.m.s.d. values presented in Table 2. The largest deviations among SCAN structures occur at the N- and Cterminal ends, which show higher flexibility than the core, and a4, which is positioned away from the dimer interface. The r.m.s.d.Figure 3. Overall structure of PEG3-SCAN. The homodimer is shown as ribbons with one subunit green, the partner purple. The Nand C- termini as well as the five a-helices of each monomer are labeled accordingly. doi:10.1371/journal.pone.0069538.gvalues for alignments with the SCAN domain dimers of Znf42 and ?Znf174 show higher variation, more than 1.0 A greater, than for the X-ray structures, because of the greater uncertainties associated with the NMR structures and that the fit involves an average of 20 conformers that represent their NMR derived structures.Residues Forming the SCAN Dimer InterfaceThe human PEG3-SCAN homodimer is held together by an extensive network of hydrogen-bonding, salt-bridge interactions and van der Waals forces. Even though the overall sequence identity among the five known SCAN structures is only 40?0 (Fig. 2), the key residues located at the dimer interface and that contribute to inter-subunit associations are conserved. TheSCAN Domain of PEGTable 2. Structure and sequence similarity between PEG3-SCAN and other SCAN domains.?R.m.s.d (A) 1.57 1.51 2.85 2.Protein name Zfp206 Znf24 Znf42 ZnfPDB codes 4E6S 3LHR 2FI2 1Y7QR.m.s.d alignment length 157 164 155Sequence identity ( ) 38 48 35These included crystal structures of Zfp206 and Znf24, and solution NMR structures of Znf42 and Znf174. R.m.s.d. calculations were carried out with PDBeFold using secondary structure matching [49] with the PEG3-SCAN dimer in the superposition. Sequence alignment was performed with ClustalW2 using residues 40?30 of the full-length PEG3 against the core of the SCAN domain, as well as 2? flanking residues, of other proteins. doi:10.1371/journal.pone.0069538.tmajority of these intermolecular contacts are formed between a1 and a2 (the N-terminal sub-domain) of one subunit and a3 on the C-terminal sub-domain of the partner. Helices a2 and a3 show the highest amino acid conservation when comparing the sequences of these known SCAN domain structures and the conserved residues cont.