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1

Golgi apparatus

- golgi conissts of a stack of membrane-bounded cisternae located b/w the endoplasmic reticulum and the cell surface - resembles a stake of hollow pancakes
-named after camillo golgi (1898)
-primary function - to modify and sort proteins for export to other organelles
-golgi is cells shipping department

2

golgi structure

bottom to top (vesicle traffic in 2 directions, transitional elements, no ribosomes)
Cis golgi network
cis cisternae
medial cisternae
trans cisternae
trans-golgi network (network of tubules , vesicles) sorting area
to: secretory vesicles (lysosomes, cell membrane)

3

each compartment (cis,medial,trans) has unique enzymes:

functional differentation of the golgi complex can be studied with the electron microscope with specific techniques that detect different enzymes:
-cis region is where sugars and proteins are phosphorylated
-the medial (middle) region have enzymes that remove carbohydrates that were added in the ER, and then add new carbohydrates
-the trans region is the area where the lysosomes are sorted. therefore, it is rich in acid phosphatase, an enzyme found in lysosome

4

golgi function

stepwise glycosylation of proteins (adding sugars):
the making of glycoproteins:
-N-linked glycoproteins
-Continued processing of N-linked sugars (from ER)
-O-linked glycoproteins

5

N-linked glycoproteins

sugars attached at the N of the asparagine amino acid (also occurs in the ER)

6

Continued processing of N-linked sugars (from ER)

-substrate is a sugar-nucleotide (activated sugar), which is transported into the Golgi from cytoplasm
-Series of glycosyltransferases, which act in a rigidly determined sequence

7

O-linked glycoproteins

-sugars attached to the O of serine of threonine amino acids
only occurs in golgi

8

why glycosylate proteins ?

-makes proteins more hydrophilic
-allows them to bind to the extracellular matrix
-reduces their susceptibility to proteases(partly b/c carbohydrates are more rigid and thereby can provide physical barriers to the core of the protein)
-provides another level of structural complexity to proteins, which can:
*give the protein a new function
*provide more specificity in its interactions with other proteins

9

where do we see an abundance of glycosylated proteins ?

on the cell's surface, where cell to cell interactions occur

10

What did John hodgson say about glycoproteins?

almost without exception, whenever two or more living cells interact in a specific way, cell surface carbohydrates will be involved

11

Golgi is a transient structure which means its ...

constantly changing

12

the following contents all move through the layers of cisternae:

-membranes including proteins, phosphoglycerides, other membrane lipids
-materials within vesicles including proteins being processed and the enzymes doing the processing

13

two models of transport through the golgi:

-cisternal maturation model
-vesicular transport model

14

cisternal maturation model

each cisterna is physically moved up the stack and changes composition.
evidence: there are some materials, including large molecules that are never seen in the vesicles

15

vesicular transport model

-materials in the interior and in the membrane are brought up to next cisternae by vesicle transport
evidence: vesicles were found to move both forward and backward (anterograde and retrograde), so they could maintain enzyme differences

16

movement of vesicles

-not random of mediated by diffusion, as people once though
-vesicles travel along microtubules
-anchored to microtubules by various proteins, including one called dynactin (only with dynein,not kinesin; kinesin attaches with help of other adaptor proteins)
-vesicles propelled along microtubules by motor proteins, such as dynein and kinesin

17

is the golgi a permanent organelle?

no
the golgi breaks up and disappears at the onset of mitosis
by telophase of mitosis, the golgi reappears
we are still uncertain how it is recreated
growing microtubules may help collect and align small vesicles which later fuse together and the plus end

18

vesicular traffic

-vesicles 60-100nm diameter, deliver soluble and membrane-bound molecules by fusion of membranes
-vesicles are covered by proteins

19

three functions of the vesicle's integral and peripheral proteins:

1. vesicle formation/budding
2. selection of vesicle contents by receptor system
3. recognition and fusing to target membrane

20

three types of vesicles

1. COP II-coated vesicles
2. COP I-coated vesicles
3. Clathrin-coated vesicles

21

COP II-coated vesicles

forward movement from ER and b/w cisternae
-anterograde movement

22

COP I-coated vesicles

retrograde movement from TGN to golgi to RER

23

Clathrin-coated vesicles

TGN to lysosomes and endocytosis

24

vesicles selectively transfer the following materials through the cell:

-contents w/in vesicles
*bulk flow, non-specific contents eg. water
*specific materials (proteins, carbohydrates, lipids) concentrated within
-specific membrane proteins

25

Some requirements of vesicular transport

-selection of contents
*receptor mediated
-production of vesicles
* by protein coat
-movement of the vesicles
*movement, tethering/docking, membrane fusion

26

selection of vesicle contents

-this requires receptors to select and carry specific contents

27

receptors are:

proteins, usually integral membrane proteins
bind other molecules (ligands) with very high specificity and very high affinity
-specificity and affinity give receptors the power to concentrate specific molecules

28

selection of vesicle contents by receptors

the coating of proteins inside a vesicle is the same as the inside of the target compartment and to the outside of the cell
ex. page 17

29

function of protein coats on vesicle exterior

-interact with the cytoplasmic tails of receptors
*ER export signals
-interact with other trans membrane proteins
-put mechanical force on membrane to form vesicles

30

three steps of protein coats on vesicle exterior

1. when coat proteins assemble at the membrane, they force the lipid bilayer to ben
2. coat proteins help gather the receptors that will carry cargo inside the vesicle
3. shortly after the vesicle pinches off, the coat falls off and the vesicle is ready to travel to destination