Exam 3 - Andreas slides READING checklist Flashcards
Going over terms not described in detail from the slides.
What is live-cell imaging?
Allows us to directly observe molecular processes in living cells.
- Involves fluorescently labeled proteins to be injected and observed.
What can fluorescence microscopy reveal?
Location within a cell of a protein present in very low concentration.
- Used on antibodies that bind with high affinity to the protein being sought.
What is the function of in vitro motility assays?
Help detect the activity of an individual protein molecule acting as a molecular motor in “real time.”
What is FRAP?
Fluorescence recovery after photobleaching
- Helps measure polymer dynamics.
Microtubules are components of what 3 things?
- Mitotic spindle of dividing cells
- Core of cilia
- Core of flagella
What is the wall of MTs made of?
Globular proteins arranged in longitudinal rows called protofilaments.
- MTs consists of 13 protofilaments aligned side by side in a circular pattern within the wall.
What kind of interactions occur between adjacent protofilaments and what is the purpose?
Noncovalent interactions play an important role in maintaining MT structure?
The plus end of a MT is terminated by what?
Beta-tubulin subunits
The minus end of a MT is terminated by what?
Alpha-tubulin subunits
What is tau?
A MAP that when expressed at abnormally high levels of phosphorylation lead to development of several fatal neurodegenerative disorders (EG. Alzheimer’s).
How are MAPs controlled?
Primarily by the addition and removal of phosphate groups from particular amino acid residues.
MTs role in plant cells
Indirect role in maintaining cell shape through their influence on the formation of the cell wall.
MT roles in cells
- Mechanical support
- Provides shape of cells
- Maintaining the internal organization of cells
What does axonal transport move?
- Material (neurotransmitter molecules) are compartmentalized within membranous vesicles in the ER and Golgi complex of the cell body and then transported down the length of the axon.
- Non-membrane bound cargo (RNAs, Ribosomes, cytoskeletal elements) are also transported down.
What is anterograde direction?
Structures and materials traveling from the cell body toward the terminals of a neuron.
What is retrograde direction?
Endocytic vesicles that form at the axon terminals and carry regulatory factors from target cells move from the synapse toward the cell body.
What classifies as the cytoskeleton?
- Microfilaments
- Intermediate filaments
- Microtubules
Kinesis and dyneins move along what?
Microtubules
Myosins move along what?
Microfilaments
What are some things that motor proteins move?
- Ribonucleoprotein particles
- Vesicles
- Mitochondira
- Lysosomes
- Chromosomes
- Other cytoskeletal filaments
What moves along intermediate filament tracks?
No motor protein known to do so.
What is different about intermediate filament tracks regarding polarity?
They are not polarized and thus would not provide directional cues to the motor proteins.
What two cycles do motor proteins employ to move along the tracks?
- Mechanical cycle (conformational changes)
2. Chemical (catalytic) cycle: provides energy through ATP hydrolysis
What is KRP?
Kinesin-related proteins
What is the responsibility of the globular heads of the kinesin molecule?
Binds to MT and acts as ATP-hydrolyzing, force-generating “engines.”
The motor portions of KRPs have related amino acid sequences, why?
This reflects their common evolutionary ancestry and similar role in moving along MTs.
Why do the tails of KRPs have diverse sequences?
This reflects the variety of cargo that these motors haul.
What are differences between myosin and kinesin?
- Kinesins are much smaller proteins.
2. Motors operate on different tracks.
Which direction does Kinesin travel?
Plus end-directed micro tubular motor.
EG. Cell body to synapse
The velocity of Kinesin is proportional to what?
The ATP concentration.
EG. Low ATP conc = slow
8 nm in length is equal to?
One tubulin dimer in a protofilament
What is processive?
Meaning that the motor protein tends to move along an individual MT for considerable distances without falling off.
Kinesins are usually well adapted for what kind of travel?
Independent, long-distance transport of small parcels of cargo.
What is different about kinesin-14?
Moves in the opposite direction (towards minus-end).
- Needs minus-minus neck regions to move this way.
- A plus-minus will always move in plus.
What is the Ncd protein of Drosophila?
A kinesin-14 protein that moves toward the minus end of the MT track.
What is different about kinesin-13?
Incapable of movement, aka MT depolymerases: bind to either end of a MT and bring about its depolymerization.
What is the protein responsible for the movement of cilia and flagella?
Dynein
What is responsible for the MT-binding site on Dyneins?
The stalks that are found at the tip of the Dynein head.
Which direction does Dynein move?
Toward the polymer’s minus end.
What are the two roles of cytoplasmic Dynein?
- A force-generating agent in positioning the spindle and moving chromosomes during mitosis.
- A minus end-directed MT motor with a role in positioning the centrosome and Golgi complex and moving organelles, vesicles, and particles through the cytoplasm.
What is the function of dynactin?
- A multisubunit protein that acts as an adaptor for cytoplasmic dynein interactions with membrane-bounded cargo.
- Regulates dynein activity
- Helps bind the motor protein to MT = increase in processivity.
What sort of cargo are dynein-driven?
- Endosomes, lysosomes,
- ER-derived vesicles heading to Golgi-complex
- RNA molecules
- HIV virus