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119 Cards in this Set
- Front
- Back
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Hoe wordt de knock out mice gemaakt? |
A DNA of a dirupted gene is introduced in ESC. Select the cells only with homologous recombination. Use therefore two selectable marker genes. Then the heterogenous knock out mice cells are inserted into a female mouse > chimera mouse. Mating of two heterogenous mices >>> knockout mice.
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WAt doet SRP en warop bindt deze?
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SRP binds to signal sequence, large ribosomal subunit.
SRP ( p54 ) aan SRPR ( alfa subunit) |
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Which proteins are the only one required for translocation ??
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SRPR,sec61 translocon
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Wat is stop transfer anchor sequence?
What type of membrane proteins have that? |
STA: both stops the passage of the pp chain through the translocon + serves as membrane anchor. Type I and III
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How do type IVa and IV proteins look like?
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a) NH3 in cytosol (+++ residues) (even = STA, odd = SA)
b) NH3 in ER ( ???) |
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What and where cleaves peptides which are then transferred to glycosilphosphatidil inositol?
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GPI transamidase and transfers the carboxyl group to GPI anchor.
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Which aa residues are substrates for glycosiltransferase?
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Asn-X-Thr, Asn-X-Ser
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What kind of glycan-tree have proteins which leave ER ??
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Man8 (GlcNac)2
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What and by which enzyme is Man8GlcNac trimmed in cis-golgi?
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endoglycosidase H (hoop ik) to Man5GlcNac2
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Which enzyme trimms mannose in transgolgi?
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Endoglycosidase D
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What do clathrin coated vesicles? Which APs are involved?
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transport from PM of TGN to endosomes>lysosomes. AP2 in PM, AP1 in TGN.
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Which sequence bind COP II vesicle sec24 subunit?
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Asp-X-Glu
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WHich sequence binds to COP I alfa and beta subunits?
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KKXX
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Welke glycosil-bomen midificaties vinden plaats in cis,med,trans golgi?
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***
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Which sequnces are incorporated in clathrine vesicles that bud from TGN?
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Try-XX-F(willekeurige hydrofobe aa)
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What is apo-/ferrotransferrin?
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***
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WHat does hrs?
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Facilitates the loading of the monoubiqutinated proteins in the buds, directed into the interior of the endosome. It also recruits ESCRT ( endosomal sorting complex required for transport) proteins.
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Which proteins there are and what is their function , like SNARE and NSF?
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ESCRT and VPS4
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WHat is TSG101?
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ubiquitine binding protein.
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What is the function of Gag?
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pinching off of the complete HIV particles.
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What do the cortex,adherence belt?
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cortex - provide support and organization
adherence belt - associated with adherence junction to provide the strenghts of the epithellium |
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Where is beta actine enriched?
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enriched in the cortex and the leading edge of the cells.
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What does alfa actin ?
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is associated with contractile structures
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What does gamma actin ?
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in stress fibers.
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What does the tropomodulin?
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Binds to the (-) end of the filament , inhibits assembly and dissassembly.
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What does gelsolin?
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Inserts itself between the subunits of the helix. it then remains bound to the + end, generating more - ends.
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Where arp2/3 binds?
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WASP and preformed actin filament.
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How do formin work?
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It has FH1,2 domains, RBD domain. RBD binds on Rho-GTP so the FH1,2 domains are exposed. FH1 domain recieves G-actin-monomeres and loads it on FH2 domain where the whole actine filament is located.
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How do WASP work?
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Similar to formin, but WASp binds op Cdc42-GTP with its RBD domain.
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What does the dystrophyn?
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Links the cortical actin network of muscle cells to a complex of membrane proteins that link the extracellular matrix.
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In which direction do myosin motor ptoteins move?
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+ direction
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What does myosin VI?
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has motility toward - end. Involved by endocytosis.
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What does myosin V?
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Binds specific organelles to be transported.
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What happens in the presence/absence of calcium in connection to tromyosin and troponin?
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In the absence of Ca2+ , TM blocks myosin interaction with F actin and the muscle is relaxed. binding of Ca2+ ions on TN triggers movement of TM to a new side on the filament > exposing the myosin binding sites on actin.
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What are the functions of these small GTP binding molecules?
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cdc42: filopodia
Rac: lamellipodia Rho: stress fibers. |
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What are the polarity of ends in the microtubules ?
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+ end - polymerization
- end - depolymerization |
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When and how do the cell microtubules are assembled?
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in interphase, from centrosome (MTOC)
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WHat does y-TURC?
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acts like template to add a-b tubulin to new microtubulli.
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Where are centrosomes,centrioles,microtubules consist of?
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microtubule: 1 protofilament , 13 repeating ab-dimers;
centriole - 9 sets of triplet mt centrosoom - one pair of centrioles. |
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Whre do the unpealing of microtubules can occur?
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When beta-GDP is not stabilized by beta-GTP
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What do colchine, taxol?
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colchine - binds tubuline dimers
taxol - binds to MTs and stabilizes them against depolymerization. |
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WHat is +Tip?
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MAP protein, which is when not phosphorylated, binds to + end of microtubules and stabilizes MTs.
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What are the differences between kinesin 13 and op18/stathmin?
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1) binds and curves the ends of MT into GDP-beta tubulin formation
2) enhances hydrolizys of GTP |
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What does kinesin 1 ?
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transport of vesicles toward + end.
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Kinesin 5?
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Pulls on antiparallel MTs
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Kinesin14?
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THe only one that moves toward - end.
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What does dynein and which protein is involved to help dynein?
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- end transport
- requires dynactin - Arp related domain of dynactin is responsible of binding cargo, p150glued - dynein binding and MT binding |
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Which 3 types of MTs are there in the mitotic spindle?
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1. Astral microtubules: orients spindle poles with axis of cell division,
2. Kinetochore MTs: transport of crhomosomes to the poles. 3. Polar MTs: pushing dublicating chromosomes apart during prophase, maintaining the structure of spindle, pushing spindle poles apart in anaphase B. |
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What does kinesin 4?
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a + end directed motor, interacts with the polar microtubules to pull the chromosomes toward the centre of spindle ( congression) prometafase.
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What does kinesin 7 ?
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Holds on the growing MTs during congression.
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What do the kinesin 5 and dynein during anaphase B ??
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Kinesin 5 associates with + end and pull the microtubules apart.
dynein - localized and anchored on the cell cortex , pulls on astral MTs and thus helps to separate the spindle poles. |
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What is integrin and what components of ECM do it bind?
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integrin is an adhesion receptor. It binds proteoglycans, collagen , fibronectin, laminin)
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What is basal lamina/ basolateral surface?
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bl- underlying basal surface ECM
bs -when basal and lateral surfaces are similar in composition. |
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What are the adherense junctions?
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cadherins, attached to actin filaments.
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What are the desmosomes ?
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Are the integrins, which are attached to IF . Responsible for cell-matrix adhesions.
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What are the desmosomes ?
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Are the desmosomal cadherins which are attached to the IF. ( cell-cell adhesions).
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Which molecules form the basement membrane?
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The basal lamina and collagen-anchroing fibrils. Sheet forming type IV collagen is a major component of the basal lamina.
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What is and does perlecan?
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Is a proteoglycan , cross-links components of the basal lamina and cell surface receptors. Perlecan has many globular repeats.
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What is basement membrane?
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Basal lamina and the immidiately adjacent collagen network form a structure called the basement membrane.
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What is the major fibrous protein in the ECM of the connective tissues?
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fibrillar collagen
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Describe type I collagen. Which type collagen coassemble with this type collagen
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Long fibers, great tension, great stretching. ( bones ) Type V and type XI collagen coassemble into fibers with type I collagen, regulating the structures and properties of the fibers.
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Describe type II collagen:
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more meshlike.
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Where are the GAGs attached?
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to glycoproteins. GAG- glycosaminoglycans.
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Wat is het verschil tussen HECT en RING families van E3 ligases?
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HECT is substaat en ubiquitine bindend eiwit.
RING is adapter eiwit voor assembly van E2-E3 complex. |
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Waarop bindt calnexin en calreticulin respectievelijk?
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calnexin bindt aan dilysine (KK) sequentie, calreticulin aan de luminale KDEL sequentie.
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Waarvoor is vacuolaire ATPase complex is noodzakelijk in lysosomen?
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voor dissociatie van M6P bevattende lysosomale pro-enzymen van de M6P.
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Wat is de oorzaak van F actine treadmilling?
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ATPase activiteit van actine
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Hoe is dynein betrokken bij de congressie?
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Kinetochoor gebonden dynein beweegt langs kinetochoor MTs in de richting van spoelfiguur pool.
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Welke region van kinesin is verantwoordelijk voor de cargo?
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De tail regio met lichte ketens van kinesin.
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Wat doet kinesin 7?
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Verbindt kinetochoor MTs met kinetochoor/chromosomen tijdens verlenging van kinetochoor MTs aan +zijde.
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Wat doet de dynein tijdens de anaphase?
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Perifeer verankerde dynien beweegt langs astrale MTs richting de centrosoom , zodat spoelfiguurpolen uit elkaar worden getrokken.
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Hoe worden a-b tubulin posttranslationeel gemodificeerd?
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Posttranslatonele modificatie van a-b tubulinnes reguleert binding van oa MT motor eiwitten assembly and severing eiwitten.
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Which functions have beta-1 integrins?
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they bind collagen and multiadhesive proteins in ECM
They also bind Ig-CAMs on the cell membrane during cell-cell adhesion. |
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Which functions have beta-2 integrins?
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They also bind Ig-CAMs on the cell membrane during cell-cell adhesion.
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What are the glycosaminoglycans?
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Glycosaminoglycanen zijn O,N linked ketens van repeterende disachariden.
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What do alfa/beta katenin ?
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Zijn de adapter eiwitten die E cadherines verbinden met circumferential belt in gepolariseerde cellen.
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Which proteins are in the ECM?
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Proteoglycans, laminins, collagen, fibronectine.
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Wat stimuleert de ATPase activiteit van Bip Atpase en wat stimuleert utiwisseling van ADP-ATP?
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Erdj1-5 en BAP respectievelijk
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Wat doen E3 ligase enzymen?
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zorgen voor de specifieke interactie met de substraat eiwitten.
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What is the function of deubiquinating enzymes?
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1)precursor processing
2)rescue from degradation 3)recycling 4)editing 5)removal of non degradative ubiquitin signal |
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Welke twee manieren zijn voor de herkenning van cargo eiwitten door COPII sec24 subunit?
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direct: binden aan asp-x-glu,LL sequence in sec24 subunit.
indirect: binden of cargos to ERGIC-53 |
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Op welke manieren herkennen COPI vesicles de cargos?
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direct: inner coat subunits KKXX sequence
indirect: KDEL receptor |
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Waaruit bestaat en wat doet de core particle?
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Deze bestaat uit 4 ringen van 7 proteases. Deze ringen hebben gated pore, waardoor eiwitten naar binnen gelust worden en afgebroken.
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Waarvoor zorgt de activator complex(regulatory particle) van proteosoom en waaruit bestaat deze?
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dat de poor open gaat. AC(RP) bestaat uit Base ( AAA ATPase - ontvouwen van eiwit, Lead - ( eiwitten die substraat herkennen ( substraat = polyubiqutineerde eiwitten), DUBS)
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Welke verschillende receptoren zijn er in de regulatory particle van de proteosoom?
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deze ubiquitine receptoren zijn: RPN10 , RPN13
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Wat is een andere manier om geubiqitineerde eiwit door reg. particle te detecteren dan door een ubiquitine receptor?
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een adaptor eiwit. Deze herkent ubiquitine domain en tegelijkertijd adaptor receptor in 19S reg.subunit.
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Wat doen Uba-Ubl shuttle eiwitten?
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targetten slecht gevouwen eiwitten naar de proteosoom core.
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Hoe zijn de high mannose, complexe, hybride N-glycanen eruit?
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High mannose: geen GlcNac op beide core mannose residue
Complexe: wel GlcNac op beide core mannose residuen. Hybride: GlcNac op een van beide core mannose residuren. |
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Welke signaal herkennen GGA en HRS?
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Ze herkennen ubiquitine op cargo eiwitten , deze dient als endosomale sorteer signaal
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Via welke vesicles/sorteersequentie worden eiwitten van TGN naar lysosomen getarget?
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AP3,clathrine vesicles: GYXXF,KKXX
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Hoe wordt uiteindelijk een functionele M6P signaal? (cis,trans-golgi).
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1.transfer van GlcNac fosfaat naar een of meerdere mannose residuen in cis golgi.
2. VErwijdering terminale GlcNac m.b.v. fosfodiesterase in TGN: vorming functionele M6P receptor. |
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Hoe binden cholesterol bevattende particles aan de LDL receptor?
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bij neutrale pH via apoB eiwit aan de LDL receptor.
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Welke internalisatiesignaal bevat LDL-R, die door clathrine adapter eiwitten wordt herkend?
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NPxY
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Welke eiwit zorgt voor de ubiquitinatie van EGFR?
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Cbl E3 ligase adapter eiwit.
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In welke coat-pits wordt clathrin gezet?
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in de clathrine coat pits.
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Wat doet gelsolin?
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knipt bestaande F-act filamenten, beschermt daarna nieuwe + uiteinden , creert daarbij nieuwe - uiteinden en bevoordeert dus turnover , remodeling van act. filamenten.
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Met behulp van welke small GTP bindende eiwit werkt formin?
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Rho-GTP
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Hoe wordt WASP geactiveerd om daarna Arp2/3 te activeren?
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Cdc42-GTP
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Wat doet myosin 1 ?
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endocytosis, membraan verankering.
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Wat doet myosine II?
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bipolaire organizatie
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wat doet myosine V?
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Transport van vesicles
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Waarme bindt myosin aan actine en de transport in welke richting bevoordeert myosin?
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ATPase head domain, + richting.
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Waarvoor dienen singlet MT?
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voor transport van vesicles and organelles
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Waarvoor dienen doublter MTs?
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cilia, flagella
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Waarvoor dienen triplet MTs?
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MTOC, zoals centriolen, basal lamina.
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Waar bevindt zich GTP cap in MTs?
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aan de + zijde, met geexposeert beta tubuline.
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Welke rol hebben N , C , M kinesins?
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1. - + transport
2. + - transport 3. destabiliseren van MT uitendien |
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Beschijf de rol van kinesin domainen?
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ATPase domain is MT bindnigsplaats, Tail domain is verantwoordelijk voor cargo interacties.
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Beschiijf de MT dynamiek in profase ( chromosome capture en congressie).
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***
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Wat doet APC bij de anaphase start?
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Zorgt voor degradatie van coherins die chromosomen rond de centromeer regio bij elkaar houden.
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BEschrijf de MT dynamiek tijdens anaphase A?
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+,- end dissasembly of kinetochrool MTs - K13.
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BEschrijf de MT dynamiek tijdens anaphase B?
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Bipolaire kinesin 5(+) end duwt polaire MTs uit elkaar > + ends dissasembly van polaire MTs. Dynein - spoel figuur separation.
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Wat doet adhesie keratin I,II?
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verbindt epitheel cellen met elkaar via desmosomen; en met basal lamina via hemidesmosomen.
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Uit welke collagen worden pezen opgebouwd?
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type I collagen + proteoglycans + type VI collagen
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Uit welke stoffen wordt kraakbeen opgebouwd?
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type II collagen fibril + type IX collagen.
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Welke intergines zijn betrokken bij cel-cel adhesie?
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type B2 integrins.
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Wat zijn de glycosaminoglycanen?
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zijn lange lineaire ser(o) gebonden ketens.
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Wat is epitheliale mesenchimale transitie?
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verlies van epitheliale karakterestieken en verkrijgen van motile mesenchimale karakterestieken.
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Waarmee gaat EMT/MET gepaard?
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1.)verlies/vorming cell matrix adhesies / cell-cell adh.
2) switch in expressie van IF 3)remodeling van actine cytoskelet 4)productie van ECM proteases (MMP) 5)verandering in cel motiliteit. |
