Lecture 13 - Organelles 3 Flashcards

1
Q

What is exocytosis?

A

Process by which newly made proteins and lipids are delivered from the ER, via the golgi, to the cell surface and cell exterior by transport vesicles that fuse with the plasma membrane

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2
Q

What are the functions of the golgi?

A
  1. Sorting and dispatching station for proteins and lipids made in ER
  2. modification of N-linked oligosaccharide chains on glycoproteins made in ER
  3. Synthesis of O-linked oligosacchardis on proteins and lipids made in ER
  4. Synthesis of glycosaminoglycan chains on core proteins of proteoglycans
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3
Q

Describe the structure of the golgi

A

Flattened membrane-bounded cisternae stacked together

Two distinct faces: cis (entrey face, adjacent to ER), trans (exit, pointing toward plasma membrane)

network of interconnected tubular and cisternal structures

Usually located near the nucleus

of golgi stacks varies per cell

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4
Q

Describe the process of protein sorting.

A

Proteins get sent through cis then trans networks and can be sent 3 ways - to the lysosome, plasma membrane, and secretory vesicle

The pathway to the plasma membrane is the DEFAULT

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5
Q

Describe the modification of N-linked oligosacchardies in the Golgi

A

An ordered series of series of sugar removals and additions carried out by glycosideases (remove sugars) and glycosyl transferases (add sugars). There is a varying amount of modification but the High Mannose form is the least modified, the “Complex” is the most modified” and the hybrid is in between the two.

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6
Q

Describe O-linked glycosylation in the Golgi

A

Covalent attachment of oligosaccharides to the OH groups of serine and threonine in proteins (o linked). Differs from synthesis of N-linked in the ER by:

Sugars are added to the proteins one at a time

Sugars are added post-translationally (by glycosyl transferases)

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7
Q

Describe the difference between proteoglycans and glycoproteins

A

Proteoglycans = small core protein, >95% carbohydrate by mass, carbs are made out of glycosaminoglycan chains made of repeating disaccharide units (6 classes of them)

Attract water to form lubricants and gels that spring back (synovial fluid)

Found in ECM and on cell surfaces

Glycoproteins - any protein with one or more covalently bound carb units that do not contain a serial repeat unti (aka no glycosaminoglycans), typically (but not always) are protein w/ little card

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8
Q

Describe the synthesis of proteoglycans in the Golgi

A

Core protein synthesized in the ER > transported to the golgi

In the golgi, glycosyl transferases act sequentially to build the 4 sugar linker region on a serine of the core protein

Repeated action of the two specific glycosyl transferases adds repeating sugars to the chain

Addt’l enzymes may modify sugars (sulfation)

Core protein can also contain N and O linked sugars

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9
Q

How are glycoproteins and proteoglycans degraded?

A

Occurs in lysosomes by hydrolases…

Endoglycosidases first remove carbohydrates from proteins

Proteases cleave protein component into amino acids which can be resued

Glycosidases act on each glycosidic bond in reverse order in which they were built (last on, first off)

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10
Q

What is the function of glycoproteins?

A

Most of the soluble and membrane proteins synthesized in the ER, including those destined for other locations, are glycoproteins.

In contrast, very few cytosolic proteins are glycosylated.

Have a ton of functions, including…. ABO blood groups, collagen, FSH, mucins, proteases,

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11
Q

What is the glycocalyx?

A

The thick layer of carbohydrate on the cell surface - help mediate wide variety of functions at the cell surface

Composed of glycolipid, glycoproteins, and proteoglycans!

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12
Q

Describe mucins

A

The most abundant macromolecules in mucus are mucins - synthesized and secreted by specialized cells within the epithelium.

Mucins are vicous glycoproteins (~80% carb by mass), most are O-linked. Often polymerize to generate hydrated gel and resistant to proteases.

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13
Q

Discuss the structures and important of ABO blood group antigens

A

They are oligosaccharide components of glycoproteins and glycolipids on surface of RBCs (extremely immungenic and important in transfusions)

The ABO gene encodes a glycosyl transferase and determines if a sugar will be added to O antigen

O antigen - encodes non-functional protein

A antigen - encodes transferase that transfers N-acetyl galactosamine (GalNAc)

B antigen - encodes transferase that transfers galactose

A and B are codominant, O is recessive

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14
Q

Describe how types O, A, B, and AB connect to each other

A

Type O - they are universal donors since they only contain O antigen, no A or B to be recognized by recipient’s immune system

Type A (AA or AO) - make GalNAc transferase, have A antigen, can only donate to A and AB. Only can accept A

Type B (BB or BO) make Gal transferase, B antigen, can only donate to B or AB. Can only receive B

Type AB - universal acceptor, have both antigens (no antibodies)

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15
Q

Describe the constitutive exocytosis pathway

A

Called the “default” pathway - all proteins get sent through this by the golgi unless they have a specific signal

Vesicles bud from trans golgi and fuse w/ plasma membrane

Operates continually in all cells

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16
Q

Describe regulated exocytosis pathway

A

Found in specialized secretory cells (those that secrete hormones or digestive enzymes)

Proteins are diverted into secretory vesicles which bud off from trans golgi and accumulate near plasma membrane

Vesicles fuse w/ plasma membrane to release contents ONLY in response to an extracellular signal

17
Q

What are the endocytic pathways?

A

Pinocytosis - uptake of fluid and small molecules in small vesicles (mediated by clathrin coats)

Continuous process in all eukaryotic cells

Phagocytosis - uptake of large particles such as bacteria in large vesicles (phagosomes)

requires receptor activation at cell surface

occurs in specialized cells (macrophages and neutrophils) - fuse w/ lysosomes and ingested material is degraded

18
Q

Discuss receptor-mediated endocytosis and an example of when this doesn’t work..

A

Classic example is of the uptake of low density lipoprotein (LDL)

Most cholesterol transported as LDL in blood

When cholesterol is needed for membrane synth, cells make LDL receptors and insert them into membrane

LDL receptors taken into cell via clathrin coated pit w/ LDL attached

Vesicles shed clathrin coat and fuse w/ endosomes (acidic, cause dissociation of LDL from receptor)

LDL transported to lysosomes and hydrolyzed to free cholesterol

LDL receptor is recycled back to cell surface

When LDL receptors are defective, cholesterol uptake is blocked and accumulates in blood

Lots of heart attacks, plaque everywhere

19
Q

What are early endosomes?

A

Located near the plasma membrane, act as the main sorting station in the endocytic pathway - acidic environment allows some receptors to release their ligands, ligands normally taken to lysosomes

20
Q

Describe late endosomes

A

Located near nucleus - endocytosed materials arrive here 5-15 minutes after uptake

Materials are ultiamtely transported to lysosoms or late endosome is converted into lysosome by import of lysosome proteins

21
Q

What are the options for receptors in early endosomes?

A
  1. Recycled back to plasma membrane
  2. Degradation - lysosome breaks that shit down ( decreases [receptor], which is known as receptor down-regulation
  3. Transcytosis - return to a different plasma membrane domain in polarized cells (ex. mother’s antibodies are transported like this to breast milk for the baby)
22
Q

Describe the structure and function of lysosomes

A

Principal sites of intracellular digestion

Contain ~40 hydrolases

Membranes contain ATP-driven proton pump that maintains the acidic pH (protects cell in case lysosome burst, the enzymes wouldn’t work at 7.2(

Membranes contain transport proteins - move digestion productions (amino acids, sugars, etc.) to cyotosol

Membrane proteins are heavily glycosylated to protect them

23
Q

How are lysosomal enzymes delivered to the lysosomes?

A

Mannose-6-phosphate is the tag that’s added to lysosomal enzymes in the ER/golgi

sorted into transport vesicles in trans golgi nextwork

24
Q

What are the 3 pathways to lysosomal degradation?

A
  1. Endocytosis (ex. LDL)
  2. Phagocytosis (ex. ingestion of bacteria)
  3. autophagy - digestion of obsolete cell parts - double membrane surrounds organelle forming autophagosome
25
Q

Describe some of the lysosomal storage diseases

A
  1. Mucopolysaccharidoses - defects in lysosomal enzymes required for degradation of glycosaminoglycans
  2. Oligosaccharidoses - “” oligosaccharides
  3. Sphingolipidoses - “” sphingolipids (ex. Tay-Sachs)
  4. Inclusion-cell disease - defect in enzyme responsible for generating M6P tag on hydrolases

hydrolases fail to target to lysosomes, so they’re secreted from the cell

Accumulation of many undigested substrates forming inclusion bodies (empty lysosomes)