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In silico modeling of human α2C-adrenoreceptor interaction with filamin-2.

Pawlowski M, Saraswathi S, Motawea HK, Chotani MA, Kloczkowski A - PLoS ONE (2014)

Bottom Line: To better understand the molecular nature and specificity of this interaction, in this study, we constructed comparative models of human α2C-AR and human filamin-2 proteins.We found that electrostatic interactions seem to play a key role in this complex formation which manifests in interactions between the C-terminal arginines of α2C-ARs (particularly R454 and R456) and negatively charged residues from filamin-2 region between residues 1979 and 2206.Phylogenetic and sequence analysis showed that these interactions have evolved in warm-blooded animals.

View Article: PubMed Central - PubMed

Affiliation: Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States of America.

ABSTRACT
Vascular smooth muscle α2C-adrenoceptors (α2C-ARs) mediate vasoconstriction of small blood vessels, especially arterioles. Studies of endogenous receptors in human arteriolar smooth muscle cells (referred to as microVSM) and transiently transfected receptors in heterologous HEK293 cells show that the α2C-ARs are perinuclear receptors that translocate to the cell surface under cellular stress and elicit a biological response. Recent studies in microVSM unraveled a crucial role of Rap1A-Rho-ROCK-F-actin pathways in receptor translocation, and identified protein-protein interaction of α2C-ARs with the actin binding protein filamin-2 as an essential step in the process. To better understand the molecular nature and specificity of this interaction, in this study, we constructed comparative models of human α2C-AR and human filamin-2 proteins. Finally, we performed in silico protein-protein docking to provide a structural platform for the investigation of human α2C-AR and filamin-2 interactions. We found that electrostatic interactions seem to play a key role in this complex formation which manifests in interactions between the C-terminal arginines of α2C-ARs (particularly R454 and R456) and negatively charged residues from filamin-2 region between residues 1979 and 2206. Phylogenetic and sequence analysis showed that these interactions have evolved in warm-blooded animals.

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The Minimum evolution tree and multiple sequence alignment of C-terminal tail of the α2C-adrenoceptor family.Panel A - proteins are indicated by the species name and the NCBI GI number. Values at the nodes indicate the statistical support for the particular branches, according to the bootstrap test. For each protein also its C-terminal sequence is presented. Sequences were aligned by MUSCLE program. Amino acids are colored according to the chemical properties of their side-chains (negatively charged: red, positively charged: blue, polar: magenta, hydrophobic: green. Only the alignment that corresponds to the C-terminal helix and flanking residues is shown. The helix was predicted by GeneSilico metaserver. Panel B - the last 14 amino acids of a2C-AR C-terminus highlighting the arginine-rich stretch (underlined). This region is conserved in mammals and in human arteriole-derived vascular smooth muscle cells (microVSM) interacts with the actin-binding protein filamin-2, shown in experimental studies to be necessary for receptor translocation to the cell surface. The numbers denote amino acids in the full-length a2C-AR polypeptide. The arrows point to amino acid residues identified by in-silico modeling to be involved in interaction with filamin-2.
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pone-0103099-g002: The Minimum evolution tree and multiple sequence alignment of C-terminal tail of the α2C-adrenoceptor family.Panel A - proteins are indicated by the species name and the NCBI GI number. Values at the nodes indicate the statistical support for the particular branches, according to the bootstrap test. For each protein also its C-terminal sequence is presented. Sequences were aligned by MUSCLE program. Amino acids are colored according to the chemical properties of their side-chains (negatively charged: red, positively charged: blue, polar: magenta, hydrophobic: green. Only the alignment that corresponds to the C-terminal helix and flanking residues is shown. The helix was predicted by GeneSilico metaserver. Panel B - the last 14 amino acids of a2C-AR C-terminus highlighting the arginine-rich stretch (underlined). This region is conserved in mammals and in human arteriole-derived vascular smooth muscle cells (microVSM) interacts with the actin-binding protein filamin-2, shown in experimental studies to be necessary for receptor translocation to the cell surface. The numbers denote amino acids in the full-length a2C-AR polypeptide. The arrows point to amino acid residues identified by in-silico modeling to be involved in interaction with filamin-2.

Mentions: Detailed analysis of the evolution of the C-terminus of α2C-adrenoceptors is presented in Figure 2, panel A. The α2C-adrenoceptors appears in nearly all sequenced Vertebrata. The α2C-adrenoceptors are divided into 5 subfamilies containing members found in: 1) Mammals, excluding Marsupials, 2) only Marsupials, 3) Birds, 4) Amphibians, 5) Reptile, and 6) Fish. Interestingly, the C-terminus of α2C-adrenoceptors is either Arginine- or Lysine-rich only in Mammals and Birds. This finding may be connected with the fact that these warm-blooded animals need systems to control temperature of the most peripheral parts of their bodies. We postulate that the α2C-adrenoceptor may be involved in the process, in which the receptor's highly positively charged C-terminal helix may be responsible for the receptor translocation.


In silico modeling of human α2C-adrenoreceptor interaction with filamin-2.

Pawlowski M, Saraswathi S, Motawea HK, Chotani MA, Kloczkowski A - PLoS ONE (2014)

The Minimum evolution tree and multiple sequence alignment of C-terminal tail of the α2C-adrenoceptor family.Panel A - proteins are indicated by the species name and the NCBI GI number. Values at the nodes indicate the statistical support for the particular branches, according to the bootstrap test. For each protein also its C-terminal sequence is presented. Sequences were aligned by MUSCLE program. Amino acids are colored according to the chemical properties of their side-chains (negatively charged: red, positively charged: blue, polar: magenta, hydrophobic: green. Only the alignment that corresponds to the C-terminal helix and flanking residues is shown. The helix was predicted by GeneSilico metaserver. Panel B - the last 14 amino acids of a2C-AR C-terminus highlighting the arginine-rich stretch (underlined). This region is conserved in mammals and in human arteriole-derived vascular smooth muscle cells (microVSM) interacts with the actin-binding protein filamin-2, shown in experimental studies to be necessary for receptor translocation to the cell surface. The numbers denote amino acids in the full-length a2C-AR polypeptide. The arrows point to amino acid residues identified by in-silico modeling to be involved in interaction with filamin-2.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4128582&req=5

pone-0103099-g002: The Minimum evolution tree and multiple sequence alignment of C-terminal tail of the α2C-adrenoceptor family.Panel A - proteins are indicated by the species name and the NCBI GI number. Values at the nodes indicate the statistical support for the particular branches, according to the bootstrap test. For each protein also its C-terminal sequence is presented. Sequences were aligned by MUSCLE program. Amino acids are colored according to the chemical properties of their side-chains (negatively charged: red, positively charged: blue, polar: magenta, hydrophobic: green. Only the alignment that corresponds to the C-terminal helix and flanking residues is shown. The helix was predicted by GeneSilico metaserver. Panel B - the last 14 amino acids of a2C-AR C-terminus highlighting the arginine-rich stretch (underlined). This region is conserved in mammals and in human arteriole-derived vascular smooth muscle cells (microVSM) interacts with the actin-binding protein filamin-2, shown in experimental studies to be necessary for receptor translocation to the cell surface. The numbers denote amino acids in the full-length a2C-AR polypeptide. The arrows point to amino acid residues identified by in-silico modeling to be involved in interaction with filamin-2.
Mentions: Detailed analysis of the evolution of the C-terminus of α2C-adrenoceptors is presented in Figure 2, panel A. The α2C-adrenoceptors appears in nearly all sequenced Vertebrata. The α2C-adrenoceptors are divided into 5 subfamilies containing members found in: 1) Mammals, excluding Marsupials, 2) only Marsupials, 3) Birds, 4) Amphibians, 5) Reptile, and 6) Fish. Interestingly, the C-terminus of α2C-adrenoceptors is either Arginine- or Lysine-rich only in Mammals and Birds. This finding may be connected with the fact that these warm-blooded animals need systems to control temperature of the most peripheral parts of their bodies. We postulate that the α2C-adrenoceptor may be involved in the process, in which the receptor's highly positively charged C-terminal helix may be responsible for the receptor translocation.

Bottom Line: To better understand the molecular nature and specificity of this interaction, in this study, we constructed comparative models of human α2C-AR and human filamin-2 proteins.We found that electrostatic interactions seem to play a key role in this complex formation which manifests in interactions between the C-terminal arginines of α2C-ARs (particularly R454 and R456) and negatively charged residues from filamin-2 region between residues 1979 and 2206.Phylogenetic and sequence analysis showed that these interactions have evolved in warm-blooded animals.

View Article: PubMed Central - PubMed

Affiliation: Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States of America.

ABSTRACT
Vascular smooth muscle α2C-adrenoceptors (α2C-ARs) mediate vasoconstriction of small blood vessels, especially arterioles. Studies of endogenous receptors in human arteriolar smooth muscle cells (referred to as microVSM) and transiently transfected receptors in heterologous HEK293 cells show that the α2C-ARs are perinuclear receptors that translocate to the cell surface under cellular stress and elicit a biological response. Recent studies in microVSM unraveled a crucial role of Rap1A-Rho-ROCK-F-actin pathways in receptor translocation, and identified protein-protein interaction of α2C-ARs with the actin binding protein filamin-2 as an essential step in the process. To better understand the molecular nature and specificity of this interaction, in this study, we constructed comparative models of human α2C-AR and human filamin-2 proteins. Finally, we performed in silico protein-protein docking to provide a structural platform for the investigation of human α2C-AR and filamin-2 interactions. We found that electrostatic interactions seem to play a key role in this complex formation which manifests in interactions between the C-terminal arginines of α2C-ARs (particularly R454 and R456) and negatively charged residues from filamin-2 region between residues 1979 and 2206. Phylogenetic and sequence analysis showed that these interactions have evolved in warm-blooded animals.

Show MeSH
Related in: MedlinePlus