Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • For Subscribers
    • For Advertisers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in

Search

  • Advanced search
Journal of Pharmacology and Experimental Therapeutics
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
Journal of Pharmacology and Experimental Therapeutics

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • For Subscribers
    • For Advertisers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Visit jpet on Facebook
  • Follow jpet on Twitter
  • Follow jpet on LinkedIn
Research ArticleArticle

Relevance of Aromatic Residues in Transmembrane Segments V to VII for Binding of Peptide and Nonpeptide Antagonists to the Human Tachykinin NK2 Receptor

A. R. Renzetti, R.-M. Catalioto, M. Criscuoli, P. Cucchi, C. Ferrer, A. Giolitti, M. Guelfi, L. Rotondaro, F. J. Warner and C. A. Maggi
Journal of Pharmacology and Experimental Therapeutics August 1999, 290 (2) 487-495;
A. R. Renzetti
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
R.-M. Catalioto
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
M. Criscuoli
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
P. Cucchi
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
C. Ferrer
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
A. Giolitti
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
M. Guelfi
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
L. Rotondaro
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
F. J. Warner
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
C. A. Maggi
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Abstract

We used membranes from Chinese hamster ovary cells stably transfected with the human tachykinin NK2 receptor, either wild-type or mutated, at four aromatic residues (His198, Tyr266, Phe270, Tyr289) located in transmembrane segments V to VII, to assess the role of these residues in the binding of natural tachykinins and peptide and nonpeptide antagonists. Three radioligands, the agonist [125I]neurokinin A (NKA), the peptide antagonist [3H]MEN 11420, and the nonpeptide antagonist [3H]SR 48968 bound to the wild-type receptor with high affinity (Kd = 2.4 nM, 0.3 nM, and 4.0 nM, respectively). Four of the six mutant receptors tested retained high affinity for at least one of the radioligands. H198A mutation abrogated the binding of NKA but not that of MEN 11420 or SR 48968 (Kd = 4.8 and 11.5 nM, respectively); Y266F mutation abrogated the binding of MEN 11420 but not that of NKA or SR 48968 (Kd = 2.8 nM and 1.2 nM, respectively); F270A mutation abrogated the binding of both NKA and MEN 11420 but not that of SR 48968 (Kd = 1.6 nM); Y289F mutation abrogated the binding of SR 48968 but not that of NKA and MEN 11420 (Kd = 2.0 and 2.9 nM, respectively). Y266A and Y289A mutations abrogated the binding of all radioligands. Among the unlabeled antagonists, the affinity of the nonpeptide GR 159897, at variance with SR 48968, resulted heavily compromised by H198A and Y266F mutations; the peptide antagonists R396 and MEN 10376 essentially followed the binding profile of NKA, but R396 showed markedly increased affinity for the Y289F mutant receptor. Taken together, these results indicate that different, partially overlapping sets of sites may be involved in the binding of agonists and diverse antagonists to the human tachykinin NK2 receptor.

Footnotes

  • Send reprint requests to: Dr. Anna Rita Renzetti, Pharmacology Department, Menarini Ricerche S.p. A., Via Rismondo 12A, I-50131 Florence, Italy.

  • Abbreviations:
    SP
    substance P
    CHO
    Chinese hamster ovary
    NKA
    neurokinin A
    NKB
    neurokinin B
    TM
    transmembrane segment
    α-MEM
    minimum essential medium α-modification
    DHFR
    dihydrofolate reductase
    hNK2R
    human tachykinin NK2receptor
    FCS
    fetal calf serum
    • Received January 29, 1999.
    • Accepted April 6, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

Log in using your username and password

Forgot your user name or password?

Pay Per Article - You may access this article (from the computer you are currently using) for 1 day for US$35.00

Regain Access - You can regain access to a recent Pay per Article purchase if your access period has not yet expired.

PreviousNext
Back to top

In this issue

Journal of Pharmacology and Experimental Therapeutics: 290 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 290, Issue 2
1 Aug 1999
  • Table of Contents
  • About the Cover
  • Index by author
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Journal of Pharmacology and Experimental Therapeutics article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Relevance of Aromatic Residues in Transmembrane Segments V to VII for Binding of Peptide and Nonpeptide Antagonists to the Human Tachykinin NK2 Receptor
(Your Name) has forwarded a page to you from Journal of Pharmacology and Experimental Therapeutics
(Your Name) thought you would be interested in this article in Journal of Pharmacology and Experimental Therapeutics.
Citation Tools
Research ArticleArticle

Relevance of Aromatic Residues in Transmembrane Segments V to VII for Binding of Peptide and Nonpeptide Antagonists to the Human Tachykinin NK2 Receptor

A. R. Renzetti, R.-M. Catalioto, M. Criscuoli, P. Cucchi, C. Ferrer, A. Giolitti, M. Guelfi, L. Rotondaro, F. J. Warner and C. A. Maggi
Journal of Pharmacology and Experimental Therapeutics August 1, 1999, 290 (2) 487-495;

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Research ArticleArticle

Relevance of Aromatic Residues in Transmembrane Segments V to VII for Binding of Peptide and Nonpeptide Antagonists to the Human Tachykinin NK2 Receptor

A. R. Renzetti, R.-M. Catalioto, M. Criscuoli, P. Cucchi, C. Ferrer, A. Giolitti, M. Guelfi, L. Rotondaro, F. J. Warner and C. A. Maggi
Journal of Pharmacology and Experimental Therapeutics August 1, 1999, 290 (2) 487-495;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • CRV431 Decreases Liver Fibrosis and Tumor Development
  • Pharmacological Characterization of Nicotine-Induced Seizures in Mice
  • Desensitization of Nicotinic Agonist-Induced [3H]γ-Aminobutyric Acid Release from Mouse Brain Synaptosomes Is Produced by Subactivating Concentrations of Agonists
Show more Article

Similar Articles

  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About JPET
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Drug Metabolism and Disposition
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0103 (Online)

Copyright © 2019 by the American Society for Pharmacology and Experimental Therapeutics