• No results found

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

N/A
N/A
Info
Downloaden
Protected

Academic year: 2021

Share "Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors"

Copied!
9
0
0

Bezig met laden.... (Bekijk nu de volledige tekst)

Download het nu ( 9 pagina )

Hele tekst

(1)

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

Peeters, M.C.

Citation

Peeters, M. C. (2011, November 17). Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors. Retrieved from

https://hdl.handle.net/1887/18092

Version: Corrected Publisher’s Version

License: Licence agreement concerning inclusion of doctoral thesis in the Institutional Repository of the University of Leiden

Downloaded from: https://hdl.handle.net/1887/18092

Note: To cite this publication please use the final published version (if applicable).

(2)

A CTIVATION OF G P ROTEIN -C OUPLED R ECEPTORS

T

HE

R

OLE OF

E

XTRACELLULAR

L

OOPS IN

A

DENOSINE

R

ECEPTOR

A

CTIVATION

(3)
(4)

A CTIVATION OF G P ROTEIN -C OUPLED R ECEPTORS

T

HE

R

OLE OF

E

XTRACELLULAR

L

OOPS IN

A

DENOSINE

R

ECEPTOR

A

CTIVATION

Proefschrift

ter verkrijging van

de graad van Doctor aan de Universiteit Leiden,

op gezag van Rector Magnificus prof. mr. P.F. van der Heijden,

volgens besluit van het College voor Promoties

te verdedigen op donderdag 17 november 2011

klokke 15.00 uur

door

Miriam Cornelia Peeters geboren te Hoevelaken

in 1982

(5)

P

ROMOTIECOMMISSIE

Promotor: Prof. Dr. A.P. IJzerman

Overige leden: Prof. Dr. T.J.C. van Berkel

Prof. Dr. J. Brouwer

Prof. Dr. G. Vriend

Prof. Dr. M. Danhof

The research described in this thesis was performed at the Division of Medicinal Chemistry of the Leiden/Amsterdam Center for Drug Research, Leiden University (Leiden, The Netherlands). The research was part of the TI-Pharma initiative “The GPCR forum for established targets” (project number D1-105).

This thesis was printed by Wöhrmann Print Service (Zutphen, The Netherlands).

(6)

To see the sakura In flower for the first time Is to experience a new sensation

Percival Lowell The soul of the Far East (1888)

Cherry Blossoms

I view knowing there must be

Something more

Tayojo (1775-1865)

(7)
(8)

C O N T E N T S

 

7

C

ONTENTS

Chapter 1 General introduction 9

Chapter 2 The importance of extracellular loops in class A GPCRs 25 for ligand recognition and receptor activation

Chapter 3 An essential role for the first extracellular loop in 45 activating the adenosine A

2B

receptor

Chapter 4 Three “hotspots” important for adenosine A

2B

receptor activation: 71 a mutational analysis of transmembrane domains 4 and 5 and the

second extracellular loop

Chapter 5 Screening for Constitutively Inactive Mutants of the 99 adenosine A

2B

receptor using S. cerevisiae

Chapter 6 The second extracellular loop of the adenosine A

1

receptor 125 plays a role in both activation and allosteric modulation

Chapter 7 General discussion, future perspectives and conclusion 151

Summary 171

Samenvatting 173

Samenvatting voor een leek 177

List of publications 181

Curriculum Vitae 183

Nawoord 185

Abbreviations 187

 

(9)

8

Referenties

Download het nu ( PDF - 9 pagina - 214.22 KB )

GERELATEERDE DOCUMENTEN

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

Glycosylated or not, EL2 is thought to play a role in receptor structure, signaling, and ligand recognition, as well as ligand binding, both orthosteric and allosteric..

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

Next, site-saturation mutagenesis was performed on positions 71 and 74 of the A 2B R. From each of the two mutagenesis sites, 100 colonies were chosen for identification.

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

In a previous study, random mutagenesis combined with a yeast screen for activating mutant receptors have been performed on two parts of the adenosine A 2B receptor in order

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

We subsequently transformed this library in the MMY24 yeast strain and started to screen at a concentration of 1 mM 3AT, a concentration at which, as a negative control, yeast

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

(B) Competition binding curves with [ 3 H]DPCPX using increasing concentrations of unlabeled CPA as displacer of wild type receptor, and mutant receptors W156A and E164A in

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

In Chapter 4, a random mutagenesis screen of the A 2B R was described containing mutations in the fragment TM4-EL2-TM5, where mutant receptors were selected that

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

A comparison with the data obtained in Chapter 4 showed that a cysteine-rich cluster in EL2 acts as a negative regulator of activation, keeping the receptor silent, whereas

Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

De gemuteerde receptoren die we identificeerden uit deze screen lieten allemaal een verminderde constitutieve activiteit zien, maar ook een verminderde activatie door de agonist..