Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • 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
  • My Cart

Search

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

Advanced Search

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

The Guanine Nucleotide Exchange Factor RasGRP Is a High -Affinity Target for Diacylglycerol and Phorbol Esters

Patricia S. Lorenzo, Maryam Beheshti, George R. Pettit, James C. Stone and Peter M. Blumberg
Molecular Pharmacology May 2000, 57 (5) 840-846;
Patricia S. Lorenzo
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Maryam Beheshti
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
George R. Pettit
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James C. Stone
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Peter M. Blumberg
  • 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

RasGRP is a recently described guanine nucleotide exchange factor (GEF) that possesses a single C1 domain homologous to that of protein kinase C (PKC). The phorbol ester [3H]phorbol 12,13-dibutyrate ([3H]PDBu) bound to this C1 domain (C1-RasGRP) with a dissociation constant of 0.58 ± 0.08 nM, similar to that observed previously for PKC. Likewise, the potent PKC activator bryostatin 1, a compound currently in clinical trials, showed high affinity binding for C1-RasGRP. Structure activity analysis using several phorbol ester analogs showed both similarities and differences in ligand selectivity compared with PKC; the differences were comparable in magnitude to those between different PKC isoforms. Similarly, the potency of the PKC inhibitor calphostin C to inhibit [3H]PDBu binding to C1-RasGRP was similar to that observed for PKC. In contrast to the relative similarities in ligand recognition, the lipid cofactor requirements differed between RasGRP and PKC. The C1 domain plus the EF-hand motif of RasGRP (C1EF-RasGRP) was markedly less dependent on acidic phospholipids than was PKCα. The differences in lipid requirements were reflected in differential ligand selectivity under conditions of limiting lipid. Despite the presence of twin EF-hand like motifs, calcium did not affect the binding of [3H]PDBu to C1EF-RasGRP. We conclude that RasGRP is a high affinity receptor for phorbol esters and diacylglycerol. RasGRP thus provides a direct link between diacylglycerol generation or phorbol ester/bryostatin treatment and Ras activation.

Footnotes

  • Send reprint requests to: Peter M. Blumberg, 37 Convent Dr., MSC 4255, Building 37, Room 3A01, Bethesda, MD 20892-4255. E-mail: blumberp{at}dc37a.nci.nih.gov

  • Abbreviations:
    GEFs
    guanine nucleotide exchange factors
    DAG
    diacylglycerol
    PKC
    protein kinase C
    [14C]DPPC
    l-α-[1-14C]dipalmitoylphosphatidylcholine
    [3H]PDBu
    [3H]phorbol 12,13-dibutyrate
    [3H]Bryo
    [26-3H]bryostatin 1
    POPC
    1-palmitoyl-2-oleoylphosphatidylcholine
    POPS
    1-palmitoyl-2-oleoylphosphatidylserine
    POPG
    1-palmitoyl-2-oleoylphosphatidylglycerol
    POPE
    1-palmitoyl-2-oleoylphosphatidylethanolamine
    POPA
    1-palmitoyl-2-oleoylphosphatidic acid
    PMA
    phorbol 12-myristate-13-acetate
    OAG
    1-oleoyl-2-acetoyl-sn-glycerol
    TRP
    transient release protein
    C1EF-RasGRP
    C1 domain plus the EF-hand motif
    LUV
    large unilamellar vesicle
    • Received September 22, 1999.
    • Accepted January 18, 2000.
  • U.S. Government
View Full Text

MolPharm articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Molecular Pharmacology: 57 (5)
Molecular Pharmacology
Vol. 57, Issue 5
1 May 2000
  • 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 Molecular Pharmacology 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.
The Guanine Nucleotide Exchange Factor RasGRP Is a High -Affinity Target for Diacylglycerol and Phorbol Esters
(Your Name) has forwarded a page to you from Molecular Pharmacology
(Your Name) thought you would be interested in this article in Molecular Pharmacology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

The Guanine Nucleotide Exchange Factor RasGRP Is a High -Affinity Target for Diacylglycerol and Phorbol Esters

Patricia S. Lorenzo, Maryam Beheshti, George R. Pettit, James C. Stone and Peter M. Blumberg
Molecular Pharmacology May 1, 2000, 57 (5) 840-846;

Citation Manager Formats

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

Share
Research ArticleArticle

The Guanine Nucleotide Exchange Factor RasGRP Is a High -Affinity Target for Diacylglycerol and Phorbol Esters

Patricia S. Lorenzo, Maryam Beheshti, George R. Pettit, James C. Stone and Peter M. Blumberg
Molecular Pharmacology May 1, 2000, 57 (5) 840-846;
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Experimental Procedures
    • Results
    • Discussion
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Fatty acid amide hydrolase in cisplatin nephrotoxicity
  • eCB Signaling System in hiPSC-Derived Neuronal Cultures
  • Benzbromarone relaxes airway smooth muscle via BK activation
Show more Article

Similar Articles

Advertisement
  • 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 Molecular Pharmacology
  • 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
  • Journal of Pharmacology and Experimental Therapeutics
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0111 (Online)

Copyright © 2023 by the American Society for Pharmacology and Experimental Therapeutics