Summary of cell biology Flashcards
Steps in intiation of protein synthesis?
Traslation begins at AUG which small ribosomal subunit searches for as TRNA binds.
Eukaryotic initiation facor (EIF-2) binds.
if correct pairing EIF-2 ATP is hydrolysed to ADP (providing headgrowth) and dissociates. Large subunit then associated.
If incorrect ATP not hydrolysed, EIF-2 not dissociated, so tRNA falls off.
Steps in translation of protein synthesis?
Incoming tRNA is associated with elongation Factor EF-TU-GTP.
If correct base pairing, promotes GTPase activity, GTP hydolysed to GDP and EF-TU dissociates and the tRNA is captured.
If incorrect tRNA no GTP-ase activity tRNA falls off.
Another Elongation Factor binds, this time close to the A site. As the GTP is hydrolysed to GDP causing a conformational change of EF-G, prmoting the movement of the tRNA into hybrid state e.g from A to P or P to E site.
Steps in termination of protein synthesis?
No tRNA for the stop codons, so releasing factor binds to A site. Hydrolysis reaction released water.
Peptidyl transferase binds H2O to the C terminus, cuasing the dissociation of the protein.
The releasing factor causes the dissociation of the ribosome.
Target sequence is cut off by?
Signal peptidase
What are two heat shock chaperones?
HSP70- allows refolding under heat stress.
HSP60- hides hydrophobic regions.
SIgnal for nuclear import? uses..
Nuclear localisation signal, ATP
Sequence of mitochondrial import?
Protein has location signal which is recognised by a receptor on the outer membrane. Bond and form a TOM complex.
This complex then looks for the TIm23 complex on inner membrane.
The two are brought closer together, protein moves through and signal peptide cleaved off.
Proteins desined for …(5) must be inserted into the ER membrane to reach their destination.
ER, Golgi, lysosome, secretory, plasma membrane.
Signal sequence for ER import is helped by?
Signal recognition particle (SRP).
Steps for ER import?
The signal sequence of the protein binds to a SRP which binds to its SRp receptor on the ER membrane.
This then finds the protein translocator and the SRP is recycled.
4 functions of the Endoplasmic Reticulum?
Protein synthesis.
Storage e.g. glycogen
Tranport
Detoxification e.g. cytochrome C.
4 more specific functions of the SER?
Synthesis of Cholesterol and phospholipids
Synthesisand storage of glycogen and glycerides
Storage of Calcium
Steroid hormone production
Two examples of Calcium release in cells?
Acinar cells-Ca release
Sarcoplasmic reticulum at neuromuscualr junction.
3 functions of the golgi apparatus?
Packaging and modify secretions for exocytosis
Renewal and modification of the PM
Delivery of materials to other organelles.
Movemnt away from the ER…
Towards…
Anterograde
Retrograde
Three coats and their functions?
Clathrin= formation of vesicles at the cell surface e.g. clathrin pits.
COP I= movement through Golgi
COP II= Promotion of vesicles from the ER to Golgi.
How do vesicles reach their correct targets?
V SNARES are encorporated into the vesicle, so even when they lsoe their coat these remain.
V SNARE interracts with T SNARE on the target cell as they are complimentary.
Form a tight junction and allows fusion with PM and import.
V SNARES recycled.
Vesicular transport model vs Cisternal maturation model?
VT: Golgi is a static structure proteins progress from one cisternae to the next by vesicle transport
CM: Golgi is dynamic, cisternae progress through the apparatus from cis to trans.
3 pathways membrane proteins delivered to cell surface?
- Signal mediated diversion to lysosomes
- signal mediated diversion to secretory vesicles ( e.g. regulated by hormones binding)
- Constitutive pathway (unregulated)
5 fates of endocytosed material?
- Degredation- by lysosomes.
- Transport back to cell surface
- Transport through
- Storage
- Recycling
Phagocytosis > ….> Pinocytosis
0.5um
What is Frustated Phagocytosis?
When two macrophages both extend psudopods round a bacterium but cant stretch round fully as other in the way
How is cholesterol synthesised in
LDL bind to LDL receptors on membrane, whihc is positioned in a clatherin pit.
Vesicle with LDL and receptor bound uncoated
Fused with early endosome.
A transport vesicle buds off to recyle the LDL receptor to the cell surface, while the now lysosome breaks down the LDL by hydrolytic enzymes to synthesise the cholesterol.
WHat is Macropinocyrosis?
When cells ruffle and non-selectively endocytose material.
Clathrin structure?
Made of a light and heavy chain, hass three legs.
4 compenents of the cytoskeleton?
Microtubules (e.g. attatch to centrosome) Microfilaments (e.g. actin)
Intermediate filaments
Associated proteins
Structure of microtubules?
Made of alpha and Beta tubulin dimers, with a GTP associated with each dimer. B exposed at growing plus end and alpha embedded in a helix.
50 KD
What does phosphorylation do to microtubules?
Phosphorylated: dynamic e.g. in mitosis
De: static.
Why does polymiserisation of microtubules happen on to the plus side? In what form?
Dimers in the GTP form can be added. Considering hydrolysis follows the polymerisation from the - to the + end, the dimers at the - end are already in the GDP state, whereas the GTP cap at the + side (which has not get got hydrolysed) can be added to or shrunk.
Function of microtubules? (3)
- Cell shape
- Movement of organelles
- Cell division
What are MAP’s?
Microtubule associated proteins:
- Motor proteins: Dynein and Kinesin
- Promote assembly: MAP2 and TAU
Structure of microfilaments?
Fires of homomeric actin, with bound ATP (minus end ADP form often when filamentous)
Can be G globular of F filamentous Actin.
42KD
What are the cell specific Actin Microfilaments?
Striated: Alpha sk or Apha c (skeletal and cardiac)
Smooth: Alpha sm and Gamma sm
Non muscle: Beta or Gamma
Example of Rapid actin polymerisation?
Acrosomal reaction in sperm force into egg.
4 types of Actin binding proteins?
- Gelsolin (caps/severs/nucleates)
- Profilin (monomer binding)
- Fimbrin (bind actin filaments together)
- Thymosin (Monomer binding also)
Structure of Intermediate filaments?
Rope like coiled coiled structures
5 intermediate filament subunit types?
Class I and II in epithelial keratins
Class III in muscle e.g. Desmin
Class IV make neurofilaments
Class V- nuclear lamins
Two microtubule motors and difference?
Dynein: Minus end directed (grows from) found in cilia or cytoplasm
Kinesin: Plus end directed and only in cytoplasm.
Two microtubule motors differences in binding to organelles?
Kinesin: Binds directly to organelles
Dynein: Binds indirectly
Roles of three main cytoskeletal components in mitosis?
Microtubules: Movement of chromosomesActin Microfilaments: Cytokinesis
Intermediate filaments: Nuclear membrane breakdown
How is the nuclear envelope broken down in mitosis?
Lamins (type of IF) holds the nuclear envelope together.
In prophase these Lamins get phosphorylated so break apart.
Dephosphylate in late telophase to reform.
What are the three microtubules that come out of the spindle poles in cell division?
Atral MT from the centrosomes
Kinetochore MT connect to kinetochore of chromosomes.
overlap microtubules maintain rigidity.
How do the daughter chromosomes move towards the poles?
Shortening of the kenetrochore Microtubules closer to the centrosome by depolymerisation
lengthening of the overlap Microtubules so centrosomes get further apart
How does cytokinesis happen?
Contractile ring made of Actin myosin constricts until is pinched off.
What is the structure of cilia and flagella?
The Axoneme:
9-2 microtubule assembly, 9 pairs of outer doublets (one complete A fibre, one incomplete B)
and one inner pair (of 2).
Has Dynein arms that contact with adjacent doublets
and inner ones.
Has radial spokes and Nexin also.
Difference between cilia and flagella?
Length:
cilia= 2-10um length 0.5 width &Whip action
Flagella= 100-200um length and 0.5 width &wave action
What are Basal bodies? structure?
Protein structures found at the base of cilia or flagella formed by centioles. They have a 9x3 microtubule structure.
How does Dynein cause shape change?
when dynein hydrolyses ATP it generates force that causes microtubules to slide over each other or bending.
Describe the sarcolemma in terms of its banding?
Actin (thin filaments) only outer section connected to the Z line is called the I band.
Region where only the thick myosin is present is the H band and both is called the A band. (dark band)
The central line is the M line, .
Calcium binds to ….. which is also bound to and moves …. out the way. This exposes the …… binding sites. The head flexes. …….. binds to …….. in order to break the cross bridge and bind further up causing the power stroke.
Troponin Tropomyosin Myosin ATP Myosin
Structure of the Myosin filament?
Coiled coil of two alpha helices with a hinge region and two head regions just after two light chains bind.
Describe Amoeboid movement?
Gel like structure with actin meshwork around the edge which can polymerise and and the gel follows.
Describe how Filapodia move?
It has focal adhesions that contain inegrins that bind to the surface of the ECM.
Actin polimerisation of plus end protudes the lamellipodia.
The end of the structure contracts and the old adhesions break and new form further along.
What are tight junctions?
(occluding junction)
Completely impermeable junction between epithelial cells. Can vary permeability for paracellular transport (in gap between cells not between).
Structure of tight junctions?
Transmembrane across the PM's sealing them. Claudin and Occludin do this. ZO proteins (zonula Occlude) link to claudin and the cytoplasmic actin filaments providing support.
What is the purpose of tight junctions?
Preserves the cells variation/polarity, segregating lipids and proteins on the PM
e.g. on the basolateral PM (outer facing) having phosphotidyl whereas on the apical glycolipid and cholesterol.
What are gap junctions?
Allows communication between cells made of clusters of transmembrane proteins forming a 2-4nm gap.
Forms pores of 2 x connexon which is made of 6 subunits of Connexin.
(Innexins in invertebrates)
What do Gap junctions let through? Regulated?
Allows passage of ions, AA, ATP, CAMP, IP3
Regulated as PH and Ca can open or close depending on levels- prevent overload.
What is he structure of an adherin junction?
Catenins bind to the actin of the first cell, and also to the cadherin dimers which span across the intracellular gap.
What is the function of an adherin junction?
Ca dependent, homophilic interactions of adhesion belts. These allow for invagination of epithelial cells where a section can be pinched off making this contractile in length.
What is Pemphigus?
Autoimmune skin blistering, removes desmoglein of the cadherin family, reduces the inegrity of skin
What are desmosomes?
Member of the cadherin family but these link to Intermediate filaments instead in cell to cell connections.
What are hemidesmosomes?
Attatches cells to the Basal lamina via integrins or intermediate filaments.
Non junctional adhesion mechanisms?
Integrins, Ig like domains, Non Ca/mg dependent
Extracellular matrix function? (5)
- Maintain tissue integrity/shape
- Proliferation- anchorage dependent growth (a surface needed for mitosis
- Differentiation- affects cell differentiation
- Migration- provides ways to move
- Microbe defence
3 components of the extracellular matrix and two exaamples for each?
- Fibrous proteins: give structure e.g collagen, elastin
- Adhesion proteins: allow cell adhesion interract with cells e.g. Fibronectin, Laminin
- Hydrated macromolecules e.g. GAG’s and Proteoglycans (protein + GAGs)
Structure of collagen?
Fibrocollagen 1 main ECM componenet.
Triple helix of 3 alpha chains
20-40 varients.
Disease associated with defects in collagen?
Ehlers- Danlos syndrome- very flexible skin but serious problems as arterials can rupture.
Collagen is extenisvely modified in the golgi how?
Hydroxylation of proline and lysine’s
Adding polysaccaride chains e.g. glycosylation of selected sugars.
……. cells make the collagen early form …. This is modified in the golgi. Self assmbly of ….chains makes the helix of ….. This is secreted out of the golgi before the …….. are cleaved into the final …. These are then bundled to make a collagen…..
Fibroblast, Pro alpha chain, 3, Procollagen, propeptides, fibril, fiber.
Elastin binds to ……… or ……. to form elastin …..?
Microfibers or fibrillin to form elastin fibres
Cross links between monomers of …… form the final elastin structure by …..
Tropoelastin by Lysyloxidase
Mutation in Elastin causes what disease? In what gene?
Marfan’s syndrome, mutation in FB1 gene (fibrilin 1 gene)
What are GAGs?
In the ECM. Charged polysaccarides that bind to proteins by covalent bonds to form protoglycans. They are all sulphated except hylauronic acid. Disaccarides 20-70 units long.
Features/ function of Hyluronic acid?
Less charged than other GAGS due to not being sulphated, which isn’t normally linked to a protein. Used for space filling, resisting compression in joints due to water attracting to it. Is secreted across the PM
What are Proteoglycans? Structure?
In the ECM. GAG’s + Protein.
The backbone is often hyaluronon with linker proteins binding to the proteoglycans . Long flexible hydrated gel.