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Synthetic peptides as structural maquettes of Angiotensin-I converting enzyme catalytic sites.

Spyranti Z, Galanis AS, Pairas G, Spyroulias GA, Manessi-Zoupa E, Cordopatis P - Bioinorg Chem Appl (2010)

Bottom Line: The rational design of synthetic peptides is proposed as an efficient strategy for the structural investigation of crucial protein domains difficult to be produced.Only after half a century since the function of ACE was first reported, was its crystal structure solved.Structural investigations of the synthetic peptides, representing the two different somatic isoform active sites, through circular dichroism and NMR experiments are reported.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, University of Patras, GR-26504, Patras, Greece.

ABSTRACT
The rational design of synthetic peptides is proposed as an efficient strategy for the structural investigation of crucial protein domains difficult to be produced. Only after half a century since the function of ACE was first reported, was its crystal structure solved. The main obstacle to be overcome for the determination of the high resolution structure was the crystallization of the highly hydrophobic transmembrane domain. Following our previous work, synthetic peptides and Zinc(II) metal ions are used to build structural maquettes of the two Zn-catalytic active sites of the ACE somatic isoform. Structural investigations of the synthetic peptides, representing the two different somatic isoform active sites, through circular dichroism and NMR experiments are reported.

No MeSH data available.


(a) Short-, medium-, and long-range connectivities. (b) Number of NOE constraints per residue (white, gray, dark gray, and black vertical bars represent, resp., intraresidue, sequential, medium-range, and long-range connectivities). (c) Schematic representation of the sequential and medium range NOEs involving HN, Hα, and Hβ protons for Zn2+-ACEC(37) (corresponds to His958-Ala994 of the human somatic form).
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fig3: (a) Short-, medium-, and long-range connectivities. (b) Number of NOE constraints per residue (white, gray, dark gray, and black vertical bars represent, resp., intraresidue, sequential, medium-range, and long-range connectivities). (c) Schematic representation of the sequential and medium range NOEs involving HN, Hα, and Hβ protons for Zn2+-ACEC(37) (corresponds to His958-Ala994 of the human somatic form).

Mentions: 1318 and 1578 NOESY cross-peaks were assigned in both dimensions for ACEC(37), and ACEN(37), respectively, in TFE aqueous solution (TFE/H20 2 : 1). The number of unique cross-peaks was 753 and 773 for ACEC(37) and ACEN(37), respectively. Their intensities were converted into upper limit distances through CALIBA [28]. The NOE-derived structural information extracted from the analysis of NOESY spectra acquired in aqueous TFE solutions under identical experimental conditions for both peptides were introduced to DYANA [29, 30] software for structure calculation (Figures 2 and 3). Structural calculations have been performed on IBM RISC6000 and xw4100/xw4200 HP Linux workstations. The family ensemble of Zn2+-ACEN(37) peptide presents root mean square deviation (RMSD) values of 0.65 ± 0.21 Å and 1.25 ± 0.24 Å for backbone and heavy atoms, respectively, and the average target function was found to be 0.39 ± 0.0164 Å2. The RMSD values of the Zn2+-ACEC(37) peptide were 0.55 ± 0.23 Å and 1.04 ± 0.27 Å for backbone and heavy atoms, respectively, and target function lies in the range 0.60  ± 4.78 × 10−2 Å2.


Synthetic peptides as structural maquettes of Angiotensin-I converting enzyme catalytic sites.

Spyranti Z, Galanis AS, Pairas G, Spyroulias GA, Manessi-Zoupa E, Cordopatis P - Bioinorg Chem Appl (2010)

(a) Short-, medium-, and long-range connectivities. (b) Number of NOE constraints per residue (white, gray, dark gray, and black vertical bars represent, resp., intraresidue, sequential, medium-range, and long-range connectivities). (c) Schematic representation of the sequential and medium range NOEs involving HN, Hα, and Hβ protons for Zn2+-ACEC(37) (corresponds to His958-Ala994 of the human somatic form).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2902127&req=5

fig3: (a) Short-, medium-, and long-range connectivities. (b) Number of NOE constraints per residue (white, gray, dark gray, and black vertical bars represent, resp., intraresidue, sequential, medium-range, and long-range connectivities). (c) Schematic representation of the sequential and medium range NOEs involving HN, Hα, and Hβ protons for Zn2+-ACEC(37) (corresponds to His958-Ala994 of the human somatic form).
Mentions: 1318 and 1578 NOESY cross-peaks were assigned in both dimensions for ACEC(37), and ACEN(37), respectively, in TFE aqueous solution (TFE/H20 2 : 1). The number of unique cross-peaks was 753 and 773 for ACEC(37) and ACEN(37), respectively. Their intensities were converted into upper limit distances through CALIBA [28]. The NOE-derived structural information extracted from the analysis of NOESY spectra acquired in aqueous TFE solutions under identical experimental conditions for both peptides were introduced to DYANA [29, 30] software for structure calculation (Figures 2 and 3). Structural calculations have been performed on IBM RISC6000 and xw4100/xw4200 HP Linux workstations. The family ensemble of Zn2+-ACEN(37) peptide presents root mean square deviation (RMSD) values of 0.65 ± 0.21 Å and 1.25 ± 0.24 Å for backbone and heavy atoms, respectively, and the average target function was found to be 0.39 ± 0.0164 Å2. The RMSD values of the Zn2+-ACEC(37) peptide were 0.55 ± 0.23 Å and 1.04 ± 0.27 Å for backbone and heavy atoms, respectively, and target function lies in the range 0.60  ± 4.78 × 10−2 Å2.

Bottom Line: The rational design of synthetic peptides is proposed as an efficient strategy for the structural investigation of crucial protein domains difficult to be produced.Only after half a century since the function of ACE was first reported, was its crystal structure solved.Structural investigations of the synthetic peptides, representing the two different somatic isoform active sites, through circular dichroism and NMR experiments are reported.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, University of Patras, GR-26504, Patras, Greece.

ABSTRACT
The rational design of synthetic peptides is proposed as an efficient strategy for the structural investigation of crucial protein domains difficult to be produced. Only after half a century since the function of ACE was first reported, was its crystal structure solved. The main obstacle to be overcome for the determination of the high resolution structure was the crystallization of the highly hydrophobic transmembrane domain. Following our previous work, synthetic peptides and Zinc(II) metal ions are used to build structural maquettes of the two Zn-catalytic active sites of the ACE somatic isoform. Structural investigations of the synthetic peptides, representing the two different somatic isoform active sites, through circular dichroism and NMR experiments are reported.

No MeSH data available.