Lecture 12 &13: endocytosis Flashcards

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1
Q

what is** endocytosis**?

A
  • a cellular process in which **substances are brought into the cell **
  • the material to be internalised is then surrounded by an area of the membrane
  • it then buds off inside the cell to form a **vesicle ** that contains the ingested material
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2
Q

Describe the different types of vesciles

A
  • vacuoles - cellular organelles that contain mostly water
  • lysosomes - vesicles that contain digestive enzymes
  • transport vesicles - vesicles that move molecules between locations
  • endosomes- specific vesicles involved in endocytosis
  • secretory vesicles - vesicles that contain materials to be secreted / released from the cell eg synaptic vesicles
  • **extracellular vesicles ** - eg ectosomes/ microvesicles - play a role in lots of biological pathways
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3
Q

Give a summary of the process of endocytosis

A
  • extracellular molecules or membrane proteins can be engulfed by the plasma membrane
  • they form vesicles which are internalised and delivered to the internal components in the cell called endosomes
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4
Q

what are endosomes? what is their function?

A
  • membrane bound organelles that are involved in regulating the trafficing of proteins, lipids etc
  • endosomes provide an environment for material to be sorted before it reaches the degradative lysosome
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5
Q

what are the 3 categories of endocytosis

A
  1. receptor mediated endocytosis
  2. phagocytosis
  3. pinocytosis
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6
Q

Describe phagocytosis

what happens from start to finsih & give eg

A
  • ‘cell eating’ - whole particles are engulfed and broken down by enzymes
  • binding of particle to receptors triggers phagocytosis, it is then taken up by the phagosome, the phagosome fuses with a lysosome
  • the fusion of the phagosome and lygosome forms a phagolysosome

example: macrophages ingesting a pathogenic organism

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7
Q

Describe pinocytosis

A
  • the cell engulfs material that has already been broken down into smaller components
  • also known as fluid endocytosis as it involves small particles from the ECF being internalised
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8
Q

what is autophagy?

A

autophagy is when cells digest their own cytoplasmic constituents (eg organelles) within lysosomes by a process related to phagocytosis
* ‘self eating’

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9
Q

how does autophagy work?

A
  • the cytoplasmic constituents are isolated from the rest of the cell within a double membraned vesicle known as an autophagosome
  • this autophagosome fuses with the lysosome and the material inside (cargo) is degraded and recycled
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10
Q

How do transport vesicles work?

A
  • transport vesicles connect one membrane compartment to another
  • they carry their cargo from the lumen of the donor compartment to the corresponding lumen of the target compartment
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11
Q

what are the 2 important functions of transport vesicles?

A
  • specifically **take up **the appropriate cargo
  • fuse only with the** correct target membranes**
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12
Q

what is located on the cytosolic surfaces of the membrane that allow the cell to segregate membranes into different subdomains ?

A

coat proteins are located on the cytosolic surface of the membranes

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13
Q

what are transport vesicles that are derived from **membrane sub-domains ** that contain coat proteins called?

A

coated vesicles

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14
Q

what are the** 2 functions** of the coat protein?

A
  • the coat protein binds to specific membrane components causing them to cluster together into a specific ‘patch’ - which makes us the vesicle membrane
  • the coat proteins then assemble into a curved basket like lettuce that causes the membrane patch to** form its spherical shape**
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15
Q

what are the 3 best studied coat proteins?

A
  1. Clarthin
  2. COPI
  3. COPII
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16
Q

what is important to note about the different coat proteins?

A
  • sometimes the same coat protein is used for more than one subcellular location ( ie membrane sub domain)
  • ie one coat protein does not belong to only 1 membrane component
  • eg clarthin is present on the plasma membrane, late endosome & early endosome
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17
Q

describe the structure of clarthrin

A
  • 3 heavy chains
  • 3 light chains
  • forms a triskelion shape
  • arranged in a 3 legged structure
18
Q

how many triskelions form a polyhedral lattice or cage around the cargo to form a vesicle ?

A

36 triskelions

19
Q

What are the N terminals interact with

A

the N terminals (ie the feet of the 3 legged structure) interact with with a inner layer of **adaptor proteins **

20
Q

what happens to the vesicle as it begins to become coated?

A
  • as the continues to form, it gradually causes the memrbane to bud (ie a little hill on the membrane) into uniformly sized vesicles
  • once the vesicle has detatched from the membrane, it then becomes uncoated
21
Q

what are adaptor proteins?

A
  • adaptor proteins form a second layer inside the clarthin cage and link it to the membrane
  • they interact with cargo receptors that bind specific soluble cargo molecules to the lumen
22
Q

What are retromers?

A
  • heteropentameric protein complex
  • they assemble on endosomes to form vesicles
23
Q

what is the function of retromer proteins?

A
  • they help recycle transmembrane receptors from the endosome to the golgi network
24
Q

under what 3 conditions does retromer assembly occur?

A
  1. only occurs if it can** bind to the cargo receptor **
  2. if it can interact with a **curved phospholipid bilayer **
  3. and if it can bind to a specific phosphoinositide
25
Q

what are phosphoinositides?

A
  • they are a family of lipids
26
Q

how do the phosphoinositides grant specifity?

A
  • both retromers and adaptor proteins in clathrin coats bind
  • they are used as molecular markers of compartment identity
  • they help to determine when and where coats will assemble in the cell
27
Q

what is dynamin?

A
  • a protein required for vesicle formation during endocytosis
  • it assembles around the neck of clathrin coated buds
28
Q

what is the function of dynamin?

A
  • it is a GTPase protein
  • causes the neck of the clathrin bud to lengthen and tighten
  • GTP hydrolysis causes neck to break
29
Q

what are rab proteins?

A
  • they are monomeric GTPases
  • G proteins
  • GTP bound rab proteins interact with the rab effector proteins on the target membrane
30
Q

what is the role of rab proteins?

A
  • they play a vital role in guiding the vesicle to the target membrane
31
Q

what are snare proteins?

A
  • one the rab proteins have guided the vesicle to the target membrane
  • snare proteins on the membrane dock the vesicles to the target membrane and they catalyse the fusion of the 2 membrane bilayers
32
Q

what are** lipid rafts**?

A

cholesterol and sphingolipid enriched microdoomains of the cell membranes

33
Q

what are vesicles that can be formed from lipid rafts called?

A

caveosomes
*

34
Q

How do caveosomes form?

A

cavolae pinch from the plasma membrane using dynamin and deliver their content into endosomal compartments called caveosomes

35
Q

what is** clathrin independent endocytosis**?

A

used to describe mechanisms of endocytosis that dont involve **clathrin or caveolin **

36
Q

what are other examples of clathrin independent endocytosis pathways?

A
  • phagocytosis and macropinocytosis
37
Q

what is an example of receptor mediated endocytosis?

A
  • transport of LDL (low density lipo-protein) that contains cholesterol into cells
38
Q

how does LDL react with it’s receptor?

A
  • the cytoplasmic region interacts with adaptin / clathrin via a motif
  • a large exoplasmic region with 40 amino acid fold binds to LDL
  • once it binds, it is then internalised by clathrin coated vesicle
39
Q

what is the first destination of uncoated vesicles?

A

endosomes

40
Q

what are the different classes of endosomes?

A
  • early endosomes (just below the plasma membrane)
  • late endosomes (close to the golgi apparatus and near nucleus)
41
Q

what are the 3 methods of sorting / receycling that occur in an endosome that contains a receptor?

A
  • recycling - the receptors return to the plasma membrane from which they originated eg LDL
  • transcytosis - specialised form of recycling in which some receptors proceed to a different domain on the plasma membrane eg antibodies
    * degradation - some receptors are delivered along with their ligands to the lysosome where they are degraded
42
Q

summary : what are the different pathways of entry into cells (endocytosis)?

A
  • phagocytosis (clathrin independent)
  • clathrin dependent endocytosis
  • caveolin dependent endocytosis
  • clathrin and caveolin independent pathways