Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V

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Standard

Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V. / Rosenkilde, Mette M; David, Ralf; Oerlecke, Ilka; Benned-Jensen, Tau; Geumann, Ulf; Beck-Sickinger, Anette G; Schwartz, Thue W.

I: Molecular Pharmacology, Bind 70, Nr. 6, 2006, s. 1892-901.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Rosenkilde, MM, David, R, Oerlecke, I, Benned-Jensen, T, Geumann, U, Beck-Sickinger, AG & Schwartz, TW 2006, 'Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V', Molecular Pharmacology, bind 70, nr. 6, s. 1892-901. https://doi.org/10.1124/mol.106.027425

APA

Rosenkilde, M. M., David, R., Oerlecke, I., Benned-Jensen, T., Geumann, U., Beck-Sickinger, A. G., & Schwartz, T. W. (2006). Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V. Molecular Pharmacology, 70(6), 1892-901. https://doi.org/10.1124/mol.106.027425

Vancouver

Rosenkilde MM, David R, Oerlecke I, Benned-Jensen T, Geumann U, Beck-Sickinger AG o.a. Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V. Molecular Pharmacology. 2006;70(6):1892-901. https://doi.org/10.1124/mol.106.027425

Author

Rosenkilde, Mette M ; David, Ralf ; Oerlecke, Ilka ; Benned-Jensen, Tau ; Geumann, Ulf ; Beck-Sickinger, Anette G ; Schwartz, Thue W. / Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V. I: Molecular Pharmacology. 2006 ; Bind 70, Nr. 6. s. 1892-901.

Bibtex

@article{5d55e710e61711ddbf70000ea68e967b,
title = "Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V",
abstract = "The extracellular part of transmembrane segment V (TM-V) is expected to be involved in the activation process of 7TM receptors, but its role is far from clear. Here, we study the highly constitutively active CXC-chemokine receptor encoded by human herpesvirus 8 (ORF74-HHV8), in which a metal ion site was introduced at the extracellular end of TM-V by substitution of two arginines at positions V:01 and V:05 with histidines [R208H; R212H]. The metal ion site conferred high-potency inverse agonist properties (EC(50), 1.7 microM) to Zn(II) in addition to agonist and allosteric enhancing properties at concentrations >10 microM. The chemokine interaction with [R208H;R212H]-ORF74 was altered compared with wild-type ORF74-HHV8 with decreased agonist (CXCL1/GROalpha) potency (84-fold), affinity (5.8- and 136-fold in competition against agonist and inverse agonist, respectively), and binding capacity (B(max); 25-fold). Zn(II) in activating concentrations (100 microM) acted as an allosteric enhancer as it increased the B(max) (7.1-fold), the potency (9.9-fold), the affinity (1.7- and 6.1-fold in competition against agonist and inverse agonist, respectively), and the efficacy (2.5-fold) of CXCL1/GROalpha. The activating properties of Zn(II) were not due to a metal ion site between the ligand and the receptor because CXCL1/GROalpha analogs in which the putative metal-ion binding residues had been substituted-[H19A] and [H34A]-acted like wild-type CXCL1/GROalpha. Based on the complex action of Zn(II) and on the chemokine interaction for [R208H;R212H]-ORF74, we conclude that the extracellular end of TM-V is important for the activation of this CXC-chemokine receptor.",
author = "Rosenkilde, {Mette M} and Ralf David and Ilka Oerlecke and Tau Benned-Jensen and Ulf Geumann and Beck-Sickinger, {Anette G} and Schwartz, {Thue W}",
note = "Keywords: Amino Acid Sequence; Base Sequence; Binding, Competitive; Circular Dichroism; Cloning, Molecular; DNA Primers; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Humans; Molecular Sequence Data; Protein Conformation; Protein Engineering; Receptors, Cell Surface",
year = "2006",
doi = "10.1124/mol.106.027425",
language = "English",
volume = "70",
pages = "1892--901",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "6",

}

RIS

TY - JOUR

T1 - Conformational constraining of inactive and active States of a seven transmembrane receptor by metal ion site engineering in the extracellular end of transmembrane segment V

AU - Rosenkilde, Mette M

AU - David, Ralf

AU - Oerlecke, Ilka

AU - Benned-Jensen, Tau

AU - Geumann, Ulf

AU - Beck-Sickinger, Anette G

AU - Schwartz, Thue W

N1 - Keywords: Amino Acid Sequence; Base Sequence; Binding, Competitive; Circular Dichroism; Cloning, Molecular; DNA Primers; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Humans; Molecular Sequence Data; Protein Conformation; Protein Engineering; Receptors, Cell Surface

PY - 2006

Y1 - 2006

N2 - The extracellular part of transmembrane segment V (TM-V) is expected to be involved in the activation process of 7TM receptors, but its role is far from clear. Here, we study the highly constitutively active CXC-chemokine receptor encoded by human herpesvirus 8 (ORF74-HHV8), in which a metal ion site was introduced at the extracellular end of TM-V by substitution of two arginines at positions V:01 and V:05 with histidines [R208H; R212H]. The metal ion site conferred high-potency inverse agonist properties (EC(50), 1.7 microM) to Zn(II) in addition to agonist and allosteric enhancing properties at concentrations >10 microM. The chemokine interaction with [R208H;R212H]-ORF74 was altered compared with wild-type ORF74-HHV8 with decreased agonist (CXCL1/GROalpha) potency (84-fold), affinity (5.8- and 136-fold in competition against agonist and inverse agonist, respectively), and binding capacity (B(max); 25-fold). Zn(II) in activating concentrations (100 microM) acted as an allosteric enhancer as it increased the B(max) (7.1-fold), the potency (9.9-fold), the affinity (1.7- and 6.1-fold in competition against agonist and inverse agonist, respectively), and the efficacy (2.5-fold) of CXCL1/GROalpha. The activating properties of Zn(II) were not due to a metal ion site between the ligand and the receptor because CXCL1/GROalpha analogs in which the putative metal-ion binding residues had been substituted-[H19A] and [H34A]-acted like wild-type CXCL1/GROalpha. Based on the complex action of Zn(II) and on the chemokine interaction for [R208H;R212H]-ORF74, we conclude that the extracellular end of TM-V is important for the activation of this CXC-chemokine receptor.

AB - The extracellular part of transmembrane segment V (TM-V) is expected to be involved in the activation process of 7TM receptors, but its role is far from clear. Here, we study the highly constitutively active CXC-chemokine receptor encoded by human herpesvirus 8 (ORF74-HHV8), in which a metal ion site was introduced at the extracellular end of TM-V by substitution of two arginines at positions V:01 and V:05 with histidines [R208H; R212H]. The metal ion site conferred high-potency inverse agonist properties (EC(50), 1.7 microM) to Zn(II) in addition to agonist and allosteric enhancing properties at concentrations >10 microM. The chemokine interaction with [R208H;R212H]-ORF74 was altered compared with wild-type ORF74-HHV8 with decreased agonist (CXCL1/GROalpha) potency (84-fold), affinity (5.8- and 136-fold in competition against agonist and inverse agonist, respectively), and binding capacity (B(max); 25-fold). Zn(II) in activating concentrations (100 microM) acted as an allosteric enhancer as it increased the B(max) (7.1-fold), the potency (9.9-fold), the affinity (1.7- and 6.1-fold in competition against agonist and inverse agonist, respectively), and the efficacy (2.5-fold) of CXCL1/GROalpha. The activating properties of Zn(II) were not due to a metal ion site between the ligand and the receptor because CXCL1/GROalpha analogs in which the putative metal-ion binding residues had been substituted-[H19A] and [H34A]-acted like wild-type CXCL1/GROalpha. Based on the complex action of Zn(II) and on the chemokine interaction for [R208H;R212H]-ORF74, we conclude that the extracellular end of TM-V is important for the activation of this CXC-chemokine receptor.

U2 - 10.1124/mol.106.027425

DO - 10.1124/mol.106.027425

M3 - Journal article

VL - 70

SP - 1892

EP - 1901

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 6

ER -

ID: 9831067