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Volumn 322, Issue 5905, 2008, Pages 1211-1217

The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist

Author keywords

[No Author keywords available]

Indexed keywords

4 [2 [7 AMINO 2 (2 FURYL) 1,2,4 TRIAZOLO[2,3 A][1,3,5]TRIAZIN 5 YLAMINO]ETHYL]PHENOL; ADENOSINE; ADENOSINE A2 RECEPTOR ANTAGONIST; CAFFEINE; DISULFIDE; GUANINE NUCLEOTIDE BINDING PROTEIN; RHODOPSIN;

EID: 56749103466     PISSN: 00368075     EISSN: 10959203     Source Type: Journal    
DOI: 10.1126/science.1164772     Document Type: Article
Times cited : (1611)

References (61)
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    • Materials and methods are available as supporting material on Science Online.
    • Materials and methods are available as supporting material on Science Online.
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    • 2A-WT refers to the wild-type construct without T4L. All constructs have a FLAG purification tag in the amino terminus and 10 histidine residues in the carboxy terminus.
    • 2A-WT refers to the wild-type construct without T4L. All constructs have a FLAG purification tag in the amino terminus and 10 histidine residues in the carboxy terminus.
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    • 2A primary sequence (accession number P29274). Note: In addition to numbering residue positions in the primary amino acid sequence, the residues have numbers in superscripts (X.YY) that indicate their position in each transmembrane helix (X, helix number, from 1 to 8), relative to the most conserved reference residue in that helix (YY). This residue is arbitrarily assigned the number 50; numbers decrease toward the N terminus and increase toward the C terminus. However, the numbering is not used in loop regions beyond residues X.20 and/or X.80 or T4L.
    • 2A primary sequence (accession number P29274). Note: In addition to numbering residue positions in the primary amino acid sequence, the residues have numbers in superscripts (X.YY) that indicate their position in each transmembrane helix (X, helix number, from 1 to 8), relative to the most conserved reference residue in that helix (YY). This residue is arbitrarily assigned the number 50; numbers decrease toward the N terminus and increase toward the C terminus. However, the numbering is not used in loop regions beyond residues X.20 and/or X.80 or T4L.
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    • In this crystal form, the crystallographic contacts are mostly driven by the T4L protein where receptor-to-lysozyme and lysozyme-to-lysozyme mainly form the lattice contacts. A relatively large receptor-to-receptor crystallographic interface (∼520 Å2) forms antiparallel receptor dimers (fig. S4, The total surface interface between receptor and T4L moieties is ∼1300 Å2, whereas lysozyme-to-lysozyme is ∼200 Å2. The largest contact interface (∼500 Å2) between receptor and T4L is noncrystallographic and is located in the cytoplasmic site, where receptor is fused to the T4L. The other receptor-to-lysozyme surface interfaces are crystallographic 260 Å2, In comparison with the previously solved β2AR-T4L fusion proteins, the T4L domain is tilted from the membrane plane and creates more surface interactions than seen in human β2AR-T4L constructs that were solved in differ
    • 2AR-T4L constructs that were solved in different space groups.
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    • The FatCat server (http://fatcat.burnham.org/) was used for structural alignment of the transmembrane helices with the rhodopsin structure 1U19 as a reference taken directly from that server: It simultaneously addresses the two major goals of flexible structure alignment; optimizing the alignment and minimizing the number of rigid-body movements (twists) around pivot points (hinges) introduced in the reference structure.
    • The FatCat server (http://fatcat.burnham.org/) was used for structural alignment of the transmembrane helices with the rhodopsin structure 1U19 as a reference taken directly from that server: "It simultaneously addresses the two major goals of flexible structure alignment; optimizing the alignment and minimizing the number of rigid-body movements (twists) around pivot points (hinges) introduced in the reference structure."
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    • We used the program Occluded Surface (OS) that calculates the occluded surface and atomic packing of protein model structures
    • We used the program Occluded Surface (OS) that calculates the occluded surface and atomic packing of protein model structures: www.csb.yale.edu/ userguides/datamanip/os/.
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    • It has been speculated that the general activation mechanism include following changes 6.47 (gauche, conformer), 6.48 (trans, conformer), 6.52 (trans, conformer) represent the active state (R*) and 6.47 (trans, conformer), 6.48 (gauche, conformer), 6.52 (gauche, conformer) represent inactive state R
    • It has been speculated that the general activation mechanism include following changes 6.47 (gauche + conformer) / 6.48 (trans - conformer) / 6.52 (trans - conformer) represent the active state (R*) and 6.47 (trans - conformer) / 6.48 (gauche + conformer) / 6.52 (gauche + conformer) represent inactive state (R).
  • 47
    • 56749149105 scopus 로고    scopus 로고
    • Basal or constitutive activity is the spontaneous production of cellular response in the absence of a ligand. An inverse agonist shifts the equilibrium toward the inactive state. An agonist shifts the conformation toward the active state. A (neutral) antagonist binds to receptors and blocks the active site but does not shift the equilibrium. A typical GPCR can dial almost any conformational equilibrium between fully inactive and fully active; therefore, an agonist or inverse agonist can be classified as partial to full. Depending on receptor and cellular environment, the nature of an inverse agonist and a truly neutral antagonist can be difficult to detect
    • Basal or constitutive activity is the spontaneous production of cellular response in the absence of a ligand. An inverse agonist shifts the equilibrium toward the inactive state. An agonist shifts the conformation toward the active state. A (neutral) antagonist binds to receptors and blocks the active site but does not shift the equilibrium. A typical GPCR can "dial" almost any conformational equilibrium between fully inactive and fully active; therefore, an agonist or inverse agonist can be classified as partial to full. Depending on receptor and cellular environment, the nature of an inverse agonist and a truly neutral antagonist can be difficult to detect.
  • 48
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    • This work was supported by the NIH Roadmap Initiative grant P50 GM073197 for technology development, Protein Structure Initiative grant U54 GM074961 for target processing, and Pfizer (R.C.S, A.P.IJ. and J.R.L. thank the Dutch Top Institute Pharma for financial support through the GPCR forum program (D1-105, The authors thank P. Kuhn for encouragement and support, J. Velasquez for help on molecular biology, T. Trinh and K. Allin for help on baculoviral expression, Q. Li and T. Mulder for biochemical and pharmacological characterization of the receptor constructs, A. Walker for assistance with manuscript preparation, and I. Wilson for scientific feedback and discussions. The authors acknowledge Y. Zheng, The Ohio State University, and M. Caffrey, University of Limerick, for the generous loan of the in meso robot [built with support from the National Institutes of Health (GM075915, the National Science Foundation (IIS0308078, and Science Foundation Ireland 02-IN1-B266, and J. Smit
    • 2A receptor.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.