Abstract/Details

The role of ligand- and cell-specific parameters in influencing G protein coupled receptor states and cell responses


2005 2005

Other formats: Order a copy

Abstract (summary)

G protein coupled receptors (GPCRs) exist in multiple dynamic states (e.g. ligand-bound, active, internalized) that influence G-protein activation and ultimately response generation. This thesis integrates experimental measurements and mathematical models of GPCR signaling to examine ligand- and cell-specific parameters and temporal characteristics of signaling important in generating cellular responses. A particular GPCR system, the N-formyl peptide receptor on human neutrophils, and more general predictions for GPCR systems are examined.

Binding of [35S]GTPγS to neutrophil membranes induced by six N-formyl peptide agonists was characterized and found to be strongly correlated with responses, suggesting that the character of N-formyl peptide-induced responses is determined early in the signal transduction cascade, at or near the receptor level. A well-characterized N-formyl peptide binding model, containing both a low-affinity and high-affinity bound receptor state, was found to predict antagonist-induced inhibition of the actin polymerization response elicited by agonist. Further, ligand-receptor binding rate constants for seven N-formyl peptide ligands were used to examine the number of receptor states required to describe the activation of neutrophils (actin polymerization and oxidant production). An additional receptor state, one not observed in kinetic binding assays, was required to account for these responses. This receptor state was interpreted as the number of low affinity bound receptors that are capable of activating G-proteins.

Finally, parameter variation and sensitivity analysis were used to study a kinetic model of general GPCR signaling and determine the ligand- and cell-specific parameters important in determining response behavior. The character of response (i.e. positive/neutral/negative agonism) was significantly influenced both by a ligand's ability to bias the receptor into active conformations and by several cell-specific parameters, including the ratio of active to inactive receptor species, the rate constant for G protein activation, and expression levels of receptors and G proteins. This latter finding is significant in that receptor and G protein expression levels are routinely varied using molecular biology techniques and can be experimentally measured. Specifically, expressing either receptor or G protein in numbers several fold above or below endogenous levels may result in system behavior inconsistent with that measured in endogenous systems.

Indexing (details)


Subject
Chemical engineering;
Biomedical research;
Biochemistry
Classification
0542: Chemical engineering
0541: Biomedical research
0487: Biochemistry
Identifier / keyword
Applied sciences; Pure sciences; Agonist inversion; G-protein coupled receptors; Ligand; N-formyl peptide receptors
Title
The role of ligand- and cell-specific parameters in influencing G protein coupled receptor states and cell responses
Author
Kinzer-Ursem, Tamara L.
Number of pages
170
Publication year
2005
Degree date
2005
School code
0127
Source
DAI-B 66/10, Dissertation Abstracts International
Place of publication
Ann Arbor
Country of publication
United States
ISBN
0542367130, 9780542367137
Advisor
Linderman, Jennifer J.
University/institution
University of Michigan
University location
United States -- Michigan
Degree
Ph.D.
Source type
Dissertations & Theses
Language
English
Document type
Dissertation/Thesis
Dissertation/thesis number
3192681
ProQuest document ID
305467017
Copyright
Database copyright ProQuest LLC; ProQuest does not claim copyright in the individual underlying works.
Document URL
http://search.proquest.com/docview/305467017
Access the complete full text

You can get the full text of this document if it is part of your institution's ProQuest subscription.

Try one of the following:

  • Connect to ProQuest through your library network and search for the document from there.
  • Request the document from your library.
  • Go to the ProQuest login page and enter a ProQuest or My Research username / password.