Signal transduction

Final Exam
 Chapter 7
CAMP .

cyclic AMP
·

Major second messenger

>
rapidly made by adenylyI cyclase
-

& diffusible
>
-

small

Quickly
>
broken
phosphodiesterase
-

down by

-
easily amplified

Adenylyl cyclase

Plasma membrane polypeptide

Catalyzes ATP .
-CAMP

Ga increases affinity of G to (2 + G
protein may facilitate interaction

G proteins /heterotrimetic G Protein) .

GTP-binding <8 complexes With Gepare activity (a subunits

GX and Go bind to PM through lipia anchors (long-chain) :

covalent .
bond
·

Activated by G protein-coupled receptor

·

Both d off States

·

on .

There's 16 Gx JGB 12 Go 1000 G possible)

·

+ . ,
- ~
form proteins (not all combinations

subfamilies
·

↳ Gs - X subunit stimulates adenyly/cyclate

4
Giv-a subunit inhibits adenylys cyclase
↳
Gq , 11 + X subunit stimulates
phospholipase B
·

structural change of Gx regulates affinity to GBf & adenylyl cyclase

Paper 6
 Purification Procedure
·

Detergent-solubilized plasma membrane Crabbit liver)

(1) DEAE-lephacel Lion exchange Column) :

According to proteins charge differences

-
Elution by gradient of Nace
-

Test each fraction for its ability to activate (

(2) Ultrogel ACA (size exclusion)

-

According to proteins' size

-
test each fraction for its ability to activate (

(3) Heptylamine-Sepharose :
hydrophobicity

(4) Hydroxyapatitie :
according to affinity to catz or

Phosphate

(5) GTP-Sepharose Protein's ability to GTP

:
bind to

(6) DEAE-Sepharose
:
according to proteins' charge

Results :

>
-
↳ F had been labeled With (32PJ-NAD+ and

Cholera toxin

& GIF has a multi subunit structure of one or more polypeptides

Chapter 8
Paper 7
Titulo del paper :
Effects of acetylcholine on the turnover of phosphoryl units in individual phospholipids
of pancreas slices and brain cortex slices.

& paper chromatogram -

phosphon, containing spots
Products
&
of mild alkaline hydrolysis of the lipids -
Ovalitatively identical .

!
Acetylcholine stimulated the incorporation of P32 into

each of the five phospholipids unequally.

Average per cent increase : 13 , 75 ,
193 1893 , 50
,

-
 Chapter 9
Calcium Ion .

Central messenger for cellular communication

·

Not made/destroyed requestered-

in a particular compartment.

Intracellular calcium concentration

·

1. "
Non-stimulated cell : <
10- M

2 .
Stimulated cell ~10 M
:

3
. Maintained by pumping out calcium from cytoplasm (to cell outside , ER , sarcoplasmic reticu

lum , muscle tissue reticulum , mitochondria) .

Calcium Signaling
Induced by inositol triphosphate (InsP3)

InsP3 binds to InSPER (receptor ; cat channel on intracellular storage membrane

·

Release of cat into cytoplasm .

calcium-binding proteins .

Remove calcium ions solution & supplying it when . Also

needed trigger
·

free from

Calcium signaling by mediating rise in free calcium

.
Paper 8
5) it only PI that is reduced by 5-HT treatment ?

>
-

Conclusion : PI breakdown can activate

calcium transport
Chapter 10
 Paper 9
Title of paper : Nitric Oxide release accounts for the biological activity of endothelium-derived relaxing factor

LEDRF)
Background :

Bradykinin is a peptide hormone known to cause blood vessels to dilate , known to induce release of EDRF from

endothelial cells

-
EDRF identity ???

Biological Assays :

The relaxations induced by down

EDRF and NO delined during passage the cascade at rates that

Were not different from each other

There was a correlation between the amount

Of NO released and the relaxation of the bioassay

tissues induced EDRF
by