Nonono Flashcards
Binding pairs for COIP
GST: glutathione
Nickle: histidine
Streptavidin: biotin
Formula for max resolution
Lamda / 2
Forward genetics
Start with a desired phenotype and mutate a bunch of genes to see what happens
Reverse genetics
Mutate gene already chose to see the new phenotype and determine original phenotype
Cell fractionation via centrifuge.
Starts slowest: first pellet : whole cells, nuclei, cytoskeleton) medium ( mitochondria lysosomes and peroxosomes) fast ( microsomes and small vessicles / everything else small)
Velocity sedimentation
Uses sucrose base gradient which allows to further seperate after cell fractions
The fast sedement are more dense and go to the bottom first ie gel
What influences rigidity
Tail length, tail saturation, temperature.
Cholesterol helps mediate
Possible movement of phospholipids
Flexión: everyday movement constant
Lateral diffusion: changing spots with neighbors
Rotation: rotation in same spot
Flip flop: rare goes from one side to other side of membrane
What makes tail longer or shorter
Long tails (saturated)
Short tails ( unsaturated)
How to tell if transmembrane domain
20+ amino acid length (will be obvious)
How is protein mobility restricted
Scaffold proteins/ other cells
Fences
Seek to avoid or not leave lipid rafts
Transmembrane / peripheral proteins
Types of trans
Single pass (1 alpha helix) n terminus can be either side
Multiple pass ( more then one alpha)
Beta baroll ( hydrophobic beta sheet makes a channel through a membrane)
Peripheral
Anchored proteins ( via amphipathic alpha helix or post translation modified group added)
Auxiliaries don’t bind directly but bind to protiens bound to the membrane
Slowest step in actin poly
Nucleation waiting to form tetromer which is heavily regulated throughout the cell
Actin is a what ase
ATPase
As actin becomes old it moves down and taken off negative end and atp is changed to ADP
Cofilin
Causes breakage in actin and speeds up minus end disassembly
Lines sides
Tropomyosin
Stabilizes actin by lining sides of the filament
Inadvertently speeds up polymerazation
Formin
Mediates and promotes actin nucleation and elongation has arms to grab and put in place
Capz
Binds to plus end and prevents additions
Tropomodulin
Caps the - end prevents subtraction treadmill
Arp 2/3
Branching of actin filaments at 70
Thymosine
Binds to g actin and prevents it from getting added on
Profilin
Binds to g actin and makes it more likely to bind to plus end
Higher order actin structures
Bundle : multiple filaments stuck together
Cross linking ( intersections of actin not caused by 2/3)
Anchoring (bound to a surface and give strength)
Branching ( caused by 2/3)
Microtubule ase
And cytoskeleton structure
GTP
Beta over alpha
Spiral
13 circle
Microtubule proteins that assist in nucleation
Gama Tusc : makes 7 copy spiral that can be built off of
Centrioles structure
Have a triplet 13 10 10
9 triplets around a sas 6 core
Cilia flagella
Doublet microtubules
13 10 ring
9 doubles around two single 13 rings inside
Centresomes mtoc
Two centrioles at 90 angle
Surrounded by pericentriolar material
Which is where gama tubulin rings like turc are which inciate nucleation
MAPS
Stabilize microtubule by binding along sides helps prevent catastrophic
Kinesin 13
Induces catastrophic and dissemble of micro tube plus side
XMAP 215
Stabilizes plus end and accelerates assembly
Y turc
Gamma tubulin ring nucleares assembly and remains on minus end
Y tusc
Small complex form together rot make 13 slots to help nucleation to occur
Can intermediate filaments form dimers
Yes yes they can
What makes intermediate filiaments replaceable
Phosphorylation
Protofilament
Monomer that come together to make diner protofibrals
Can be added on at both sides
Keratins
Epithelium
IF
Give strength
Neuro filaments
Help the axon grow
Streagth en crosssection
Nestin
Also in neuron
Helps to maintain width
IF
Lamins
Makes a mesh framework in nucleus
IF
Vimentin
Involved in cell signals / tight junction
IF
Dynein
Processive
Minus end
MT
Kinesin
Processive
Plus end
MT
Myosin 1
Non processive
Plus ended
(Vessicles transport)
Myosin 2
(Non processive )
Plus end
Muscle contraction
Myosin 5
Processive plus end motor
Involved in vessicles trans
Myosin 6
Processive minus end
Vessicle trans
Hemidesmosmes
Use integrins
Like intermediate filaments
Via integrin dimers and plectin adapters
Cell to matrix junction
Laminin and collagen
Focal adhesión
Use integrins
Link integrin dimers to actin through talin and vinculin
Cell to matrix
Desmosomes
Uses cadherins
Bound to Inter fill via plaques
Cell to cell jubction laminin and collagen
Adherins junctions
Use cadherins bound to actin via alpha beta catenins
And vinculin (adapting protien)
Cell to cell jubction)
How are integrins activated
RAP1 which is a GTPase
Binds RIAM and recruits adapter proteins
Active not holding hands
Inactive holding hands
Cadherin activation
Adhering that bind in the presence of Ca
Most are homophonic
Tight junctions
Made up of Claudins and occludins
Make a ring around the cell and make a tight seal
Don’t bind to cytoskeleton
And therefore can’t help tissue strength but instead fill gap
Who do snoRNPs do
They methylate and pseudouridylation something
Order of 45s maturation
Chemistry change by snoRna and then cleaved into 3 parts
28s 5.8s to 60s
18s to 40s
5 s goes to 60 but made elsewhere
Space between 2 lipid layers of nucleus
Peri nuclear space
Lumen of ER
The nucleolis location and structure
Mostly euchromatin with ribosome portiens and chaperone proteins
It is fused with nuclear membrane
Linc complex
Outside Nesprin2: which interacts with cytoskeleton
Emerin sun : forms link From inside to outside and incudes nuclear lamina which binds transcription depressors
Enron and sun also bind BAF
BAF binds to chromatin
How do lamins affect transcription what proteins are close by
They form chromosome scaffold to hold dna at specific places which impacts the amount of transcription that occurs in a certain region. Holds lamin A B1 B2 C and forms a network and is part of linc complex
