Regulation of membrane transporters by kinases [Elektronische Ressource] / von Manzar Banoo Shojaeifard

Physio

Sujets

• Biologie

Informations

• Publié par: eberhard_karls_universitat_tubingen
• Publié le: 01 janvier 2006
• Nombre de lectures: 24
• Langue: Deutsch
• Poids de l'ouvrage: 1 Mo

Extrait

Regulation of membrane transporters by kinases




der Fakultät für Biologie
der Eberhard Karls Universität Tübingen

zur Erlangung des Grades eines Doktors
der Naturwissenschaften


von

Manzar Banoo Shojaeifard
aus Shiraz / Iran
vorgelegte
D i s s e r t a t i o n
2006























Tag der mündlichen Prüfung: 2.11.2006
Dekan : Prof. Dr. F. Schöffl
1. Berichterstatter: Prof. Dr. med. Florian Lang
2. Berichterstatter: Prof. Dr. rer. nat. F. Götz





























Aus dem Institut für Physiologie
der Eberhard-karis-universität Tübingen
Abteilung Physiologie 1
Leiter : Prof. Dr. med. Florian Lang



Regulation of membrane transporters by kinases



Fakultät für Biologie
der Eberhard Karls Universität Tübingen

zur Erlangung des Grades eines Doktors
der Naturwissenschaften


von

Manzar Banoo Shojaeifard
From Islamic Republic of Iran

vorgelegte
D i s s e r t a t i o n
2006





CONTENT_______________________________________________________________

List of Abbreviations......................................................................................................i
INTRODUCTION.............................................................................................. Page
I-........ Electrophysiology.............................................................................................. 1
I- A.....Voltage Clamp :................................................................................................. 1
I- A. 1.......Motivation...................................................................................... ............... 1
I- A. 2.......Background.....................................................................................................1
I- A. 2. a..... Theoretical background of voltage clamp...... ............................................1
I- A. 2. b......The ideal voltage clamp...............................................................................1
I- A. 2. c.....The Real voltage clamp.................................................................................2
I- A. 2. d....The voltage clamp with two electrodes (TEVC)............................................3

I- B... Control of membrane potential........................................................................5

I- C....Microelectrodes & micropipettes ....................................................................6

I- D....Xenopus oocytes as expression model for heterologous membrane proteins7

I- E ...Role of foreign RNA...........................................................................................9

I- F.... Transporter...........................................................................................10
I- F.. 1.... SLC ( Solute Carrier ) series........................................................................10
I- F.. 2.. Creatine Transporter CreaT (SLC6A8).......................................................13
Sodium-Phosphate Cotransporter (NaPi-2b).....................................................14
I- F.. 3.....

I- G. ..Substrates .........................................................................................................16
I- G. 1... ....Creatine (Cr ) .............................................................................................16
I- G. 2... ...Sodium-Phosphate......................................................................................18

I- H. Protein Kinases:................................................................................................18
I- H. 1....SGK & Isomers.............................................................................................18
I- H. 2... Protein Kinas B (PKB/Akt)..........................................................................19
I- H. 3... Neuronal cell expressed developmentally downregulated 4-2(Nedd4-2).....20
I- H. 4....Rapamycin (C H NO ).............................................................................21 51 79 13
I- H. 5....The Mammalian Target of Rapamycin (mTOR)...........................................23

MATHERIALS & METHODS
II-...... List of Laboratory Equipment.......................................................................26

? -.... List of Materials and Chemicals.....................................................................27

IV-... Solutions used for enzymatic defolliculation, storage.................................. 28

V-.... Aim of the present study...................................................................................28
V- A. Experimental setup............................................................................................29
V- A. 1. Prepration of oocytes.....................................................................................30
V- A. 2. cRNA Injection..............................................................................................31
V- A. 3. Preparation of microelectrodes......................................................................32

CONTENT_______________________________________________________________

V- B. Methods:...........................................................................................................33
V- B. 1. Constructs.....................................................................................................33
V- B. 2. Electrophysiology.........................................................................................33
V- B. 3. data Analysis.................................................................................................33

RESULTS.................................................................................................34
VI- A Stimulation of the creatine transporter SLC6A8:..............................................34
VI- A. 1 by the protein kinases SGK1 and SGK...................................................34
VI- A. 2 by the protein kinase mTOR.........................................................................38
VI- B Stimulation of the intestinal phosphate transporter SLC34A2:..........................42
VI- B. 1 by the protein kinase mTOR...........................................................................42

DISCUSSION...........................................................................................47
VII- A... :...........................................47
VII- A. 1... by the protein kinases SGK1 and SGK3...............................................47
VII- A. 2....by the protein kinase mTOR.......................................................................48
VII- B... Stimulation of the intestinal phosphate transporter SLC34A2:......................48
VII- B. 1... .......................................................................48
Summary…............................. ....................................................................................50
Zusammen…………………. ......................................................................................51

REFERENCES ......................................................................................................52
Acknowledgments.......................................................................................................68
Curriculum Vitae........................................................................................................70
List of Publications.....................................................................................................73




































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