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MCO test 3 Flashcards

1
Q

membrane function

A

semipermeable barrier
detects and interprets changes in extracellular environment
anchorage for proteins and cytoskeleton

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2
Q

lipid structure

A

diverse
defined by hydrophobicity rather than structure
soluble in organic solvents

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3
Q

phospholipids

A

amphiphilic/amphiphatic
polar = hydrophillic
fatty acids are hydrophobic

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4
Q

ampiphatic

A

both properties

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5
Q

membrane typically contains _ lipids

A

109 lipids

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6
Q

_____ lipid molecules per __ of bilayer

A

5 x 10^6 per 1 micrometer squared

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7
Q

internal membrane modifications done in ER only found on ___ side

A

exo

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8
Q

micelle liposome

A

hydrophobic

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9
Q

fatty acids

A

terminate with carboxylic acid group
long hydrocarbons (14-24C) 16/18 most common

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10
Q

saturated fatty acids means no

A

double carbon bond
so straight chain

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11
Q

C:D

A

number of carbons to numbers of double bonds

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12
Q

C=C

A

introduces kink in chain causes irregular packing (leading to lower melting point) OR more fluidity

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13
Q

essential fatty acids must be

A

Obtained through diet

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14
Q

smedley-maclean

A

Arachnoid acid 20:4
synthesised from clinic acid precursor for eicosanoids

also important for phospholipid bilayer
plays important role in inflammation
(diagram)

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15
Q

phospholipids can be modified by

A

ester bonds between phosphate groups

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16
Q

common head groups

A

have biological functions
phosphaldyl choline and phoshatidlynositol can be cleaved inositol and choline are important signalling molecules

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17
Q

PLC is activated process

A

1)signal transmits to phospholipase (PLC) only some pathways depend on phosphatidylserine other membrane proteins important
2)inositol phosphate is cleaved and transmits signal into cell

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18
Q

sphigomyelin

A

sphingosine with a fatty acid and hydrocarbon chain andcholien attached

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19
Q

choline

A

in myelinated axons signal transduction apoptosis

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20
Q

glycolipids

A

-sugar containing lipids instead of P group
-can be more than one sugar unit
-dereived from sphingosine not glycerol
-sugar always on outside of cell

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21
Q

cholesterol

A

-modified steroid
-cholesterol 4 hydrocarbons only steroid ring is in membrane

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22
Q

glycolipids role

A

for immune responses
cell recognition
attachments lipids

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23
Q

importance of membrane fluidity

A

-lipids diffuse laterally
-proteins not involved in anchoring also diffuse
-proteins need to transmit signals
-transport across by diffusion or via transporter
-vesicles need to bud off and fuse

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24
Q

measuring rate of lateral diffusion in membrane

A

-membrane with flurophobes
-intense light bleaches flurophobes
-rate of diffusion of flurophobes can be measured

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25
Q

how fluid is the membrane

A

-biological membranes have constant movement within the bilayer
-rate of membrane lipid movement can be measured

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26
Q

lateral diffusion per second

A

2 micrometre Per second (a length of the mammalian cell)

transverse diffusion flip flop once every 3 days (rare)

proteins are similar but generally move slower

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27
Q

membrane fluidity temperature

A

too fluid = membrane disordered too much permeability
too solid = gel slows down movement too much

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28
Q

lipid molecules move faster as temperature increases so membrane becomes more

A

permeable

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29
Q

increases fluidity

A

-unsatured lipids gives kinks
-short chains allow fewer interactions between lipids
-high temperature

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30
Q

if you cannot intrinsically reg your temp then you must adapt to surroundings

A

-organisms regulate their lipid composition
-short unsaturated fatty acids predominate at low temp
-long saturated fatty acids predominate at high temps

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31
Q

plants and their response to heat

A

-plants have sensors in the plasma membrane that detect changes in fluidity
-fluidity increases indicates temp is increasing
-allows the plant to prepare for heat stress

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32
Q

no cholesterol in

A

plants and bacteria

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33
Q

ethanol increases

A

membrane fludity

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34
Q

lipid bilayers are

A

asymmetric
-teh two layers have different lipid composition
-transverse diffusion (flip-flop) once every 3 days (rare)
-proteins known as phospholipid translators (flippases) catalyse the flip-flop event to maintain phospholipids in the correct monolayer

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35
Q

types of integration membrane proteins

A

-single span hydrophobic alpha helix either C or N terminal can be intracellular
-multi-spanning containing alpha helices , 7 transmembrane helix protein a big family but can have ore or fewer helices
-beta barrel protein forming a pore

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36
Q

membrane topology =

A

= arrangement relative to membrane this does not change

maintained by hydrophobic and electrostatic interactions

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37
Q

positively charged amino acids interact with

A

negatively charged lipid head groups

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38
Q

integral membrane proteins structure

A

-distinguish between transmembrane cytoplasmic and extra-cellular parts
-loops can form binding sites there may be entire protein domains with extra or intra cellularly

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39
Q

ICAM has several extracellular domains

A

ICAM is involved in cell adhesion
-expressed in cells of the immune system and endothelial cells
-up-regulated during inflammation
-it has 5 extracellular immunoglobulin domains

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40
Q

ICAM has

A

-5 extracellular immunglobin domains
-single transmembrane spanning helix
-short cytoplasmic tail

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41
Q

porins

A

-forms a barrel shaped structure with a pore in the centre
-8 beta strands

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42
Q

peripheral membrane proteins

A

do not interact with the hydrophobic core of the membrane
-can be cytoplasmic or ectoplasmic
-interact with lipid head groups and integral membrane proteins
-interactions are non-covalent
-electrostatic iteraction, H bonds and van Der Waals bonds

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43
Q

palmitylation

A

one of many types of lipid anchors

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44
Q

peripheral membrane proteins

A

proteins coanchored to the membrane through hydrocarbon groups

the protein is covalently attached to a hydrocarbon group

they hydrophobic hydrocarbon group inserts into the lipid bilayer

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45
Q

ankyrin and spectrin

A

-spectrin cytoskeleton protein creating a scaffold on the intra-cellular side of membrane
-ankyrin binds to several integral membrane proteins AND to spectrin
-maintain plasma membrane integrity via the spectrin-actin based cytoskeletal structure

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46
Q

cells are covered in carbs

A

-only found in ectoplasmic side of membranes
-attached to both lipids (glycolipids) and proteins (glycoproteins)
-the glycocalyx is a network of glycoproteins with mucus like consistency

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47
Q

carbs on cells role

A

-physical barrier (protects against viruses and bacteria)
-mechanosensing
-possible roles in cell shape

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48
Q

most protein have at least one carb unit few (___) lipids have carb units
exist as either oligosaccharide chains or single sugar residues

A

10%

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49
Q

glycoproteins usually have

A

oligiosaccharide chains

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50
Q

lycolipids usually have

A

single sugar residues

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51
Q

membrane carbs function

A

-cell recognition communication and adhesion
-this is especially important in immune responses
-distinguishing self and non-self, infection and transplantation

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52
Q

glycans

A

are on the outside of membranes and attached to either proteins or lipids

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53
Q

what can cross lipid bilayers

A

-small hydrophobic molecules
-small uncharged polar molecules
-water
-large uncharged polar molecules
-charged ions
-charged polar molecules

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54
Q

transport by simple diffusion

A

-solute must be hydrophobic to dissolve in membrane
-rate of transport depend on size and hydrophobicity
-rate and direction depend on concentration gradient (transport continues to a dynamic equilibrium)

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55
Q

channels

A

gated (voltage, ligand mechanical)

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56
Q

carriers

A

-permeases, transporters or carriers

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57
Q

three classes of active transporters

A

-P type pumps, phosphorylate themselves during transportation cycle (ion gradients Na, K and Ca)
-F-type pumps - work in reverse using proton gradients to synthesise ATP
-ABC transporters - pump small molecules as opposed to ions

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58
Q

1 active transporters

A

conformational change in membrane

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59
Q

P-type pump

A

lysosome need low PH for hydrolytic enzymes to be activated

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60
Q

transport of several ions makes stomach acid

A

1)CO2 diffuse in from blood
2)Combines with water to form carbonic acid catalysed by carbonic anhydrase
3)bicarbonate exchanged for CL-
4)H+ transported by P pump
5)CL- enters the lumen via a CL- channel

61
Q

HeLa human cervical cancer cellist tissue culture Henrietta lacks died 1951 aged 31

A

-it is easy to spot daughter cell pairs it is uncommon to see to see a dividing or a dying cell even in relatively rapidly growing tissues (HeLa cells divide once every 24 hrs)

62
Q

1665 Hooke published micrographia and described living tissues being composed of

A

these pores or cells

he later confirmed the observations of Antoine van Leeuwenhoek that there were single cell animalcules

63
Q

1875 Mayzel

A

cell division was described carefully by the polish histoligist

-he showed that salamander embryo cells took up aniline dyes that stained condensed structures in the nucleus (chromosomes or coloured bodies)

64
Q

1870s phases of mitosis and cytokinesis in a mammalian cell

A

-the still images of the early micrographers could be put into a sequence of phases that described the life history of cell

65
Q

interphase (G2) FISH the technique

A

-cells are grown on glass slides, fixed and permeabilised with detergent

-incubated with fluorescent oligonucleotide probes specific for individual chromosomes

66
Q

interphase (G2) primers

A

-they hybridise with their targets the the chromosomes become painted
,
-interphase chromosomes occupy their own territories. They dispersed structure allows access of transcription factors to the DNA: the cells are actively making RNA and proteins

67
Q

micro tubular spindle fibres grow from the region adjacent to the centrosomes : some extend pole to pole and

others attach to chromatids at

A

kinetochores

68
Q

1983 Tim hunt

A

identified the first controller
process about methionine added at t = 0 **

69
Q

cyclin is a controller

A

concentrations rise and fall

70
Q

cell cycle progression is controlled by heterodimeric complexes each comprising

A

regulatory cyclin that selects the targets and catalytic cyclin dependent kinase that phosphorylates a target protein

71
Q

multiple cyclins and multiple CDKs

A

the important principle is that there are G1 complexes which act as G1 controllers

cyclin CDK complexes phosphorylate their targets which prepare the cell for S phase and promote the expression pf S phase cyclin

72
Q

S phase cyclin CDK complex phosphorylates

A

its target which control chromosomes replication

73
Q

The S phase cyclin CDK

A

complex phosphorylates its target which control chromosomes replication

74
Q

how is spindle formation activated

A

G2/M cyclin complexes phosphorylate their targets

75
Q

one CDk adapted by different cyclins led to the idea

A

if this was an evolutionary intermediate and investigated by NURSE in 1996

76
Q

marine alga O. tauri a photosynthetic machine had

A

genome sequenced 2004-2006 and there are two cell cylcle cyclins only

the G1 D and G2B

77
Q

fission yeast Sz

A

1996 Genet a quantitive model for the ccdc control of S phase and mitosis in fission yeast

2010 found answer that the driving of the cell cycle with a minimal CDK control network

78
Q

looking at Sz pome survive with one CDK

A

1)the CD13 and CDC2 genes were fused with an in frame linker

2)expressed in Sz. Pombe

3)all other cyclins and CDKs were deleted one by one checking for viability each time

implications:a primoridial cyclin and a primordial CDK were sufficient to control the cell cycle and gene duplication of cyclin or of the CDK allowed new combinations to perform at different stages ion the cell cycle;e

resulting in complexity seen in modern oranisms

79
Q

cells can leave the cell cycle and enter a

A

quiescent phase

80
Q

What can a cell do?

A

-not too much room for division uncontrolled proliferation = cancer

81
Q

If these G0 arrested cells then re-enter the cell cycle a

A

tumour may reform

82
Q

Activation of growth factor receptors

A

they start at the plasma membrane by binding of specific ligands (EGF epidermal growth factor)

83
Q

Activated EGFR is bound by adaptor molecules that

A

recruit and activate the cytosolic membrane bound ras

Biological significance the signal has been transducer from the extracellular side of the plasma membrane to the intracellular side of the membrane

84
Q

Signal transduction via an enzyme cascade

A

Amplifies the signal

85
Q

growth factor summary

A

highly conserved signal transduction pathway. Stimulation of growth factor receptors results in Raw recruitment, signal transduction from Raf to MAPK and expression of G1 cyclins and CDKs that return to G0 cell to G1

86
Q

what can go wrong

A

causing unregulated prolifearation

87
Q

tumour suppressor genes arrest the cell cycle in

A

G1 some in G2

88
Q

they suppress G1 by

A

slowing down entry into S phase

giving time for DNA to repair following mitosis

key regulator is pRb(retinoblastoma protein)

89
Q

how does pRb work?

A

S phase proteins are under the control of E2F transcription factors in G1 pRb binds E2Fs and inactivates them so the cell cannot make S phase proteins until

the G1 cyclin /CDKs have accumulated enough to phosphorylate Rb and inactivate it allowing S phase proteins to be made

90
Q

p53 another tumour suppressor protein

A

loss of protein creates genome instability resulting in aneuploidy

91
Q

TP53

A

tumour suppression is severely reduced if damaged

associated with 50% of human cancers

92
Q

DNA damage, hypoxia and other stresses

A

cell cycle arrest, DNA repair and cell cycle restart

or apoptosis death of damaged cells

cellular and genetic stability

93
Q

human papilloma virus inhibits

A

p53 activity and others* check lecture 27 for specifics

94
Q

necrosis

A

cells that die through tissue damage exhibit different morphological changes

dying cells swell and burst and intracellular contents are released into extracellular milieu
causing inflammation

95
Q

2002 Nobel prize Brenner, Horvitz and Sulston

A

for their discoveries of genetic regulation of organs development and programmed cell death

96
Q

apoptosis

A

dying cells shrink

97
Q

capase

A

cleaves target at just the C terminal leading to cell death

normally kept inactive by trophic signals from neighbouroring cells
therefore apoptosis is our natural state

98
Q

internal triggers for apoptosis

A

recognition of irreparable DNA damage by the p53 pathway
DNA damage from radiation, chemo mistakes in replication and lack of trophic signalling

99
Q

external triggers for apoptosis

A

-recognition of stress
-heat, radiation and starvation
can damage cell membranes leading to apoptosis

100
Q

developmental triggers for apoptosis

A

apoptotic programs remove the webbing between our fingers during foetal development and remove a number of neurons

101
Q

GFP tagged proteins

A

help see localised sub cellular structures

102
Q

how do proteins reach their destination

A

sorting signals

they are part of the protein
-short peptides at N or C terminal = can be removed after use or kept on if needed again
-3 dimensional domains (secondary tertiary structure) = for transport like lysososomes
-other molecules attached to the protein like post-translational modifications of sugars and lipids

103
Q

what happens to sorting signals

A

-they are recognised by specific receptors
-which in turn trigger transfer of client protein to correct destination
-every organelle uses different receptors and different sorting processes
-if any of this processes goes wrong the cell is in trouble

104
Q

models of protein transport-gated transport

A

import into and export out of the nucleus

105
Q

models of protein transport-transmembrane transport

A

protein import into ER and mitochondria uses transient translocation channels in the correct membrane

106
Q

models of protein transport-vesicular transport

A

secretion along the organelles of the secretory pathway we will cover targeting to lysosomes

107
Q

gated transport into the nucleus

A

proteins and other macromolecules move between the cytoplasm and nucleus via large aqueous nuclear pore complexes

108
Q

a mammalian nuclear envelope contains 3000-4000 nuclear pore complexes

A

-side view and face-on EMs of nuclear envelope showing the distinct 8 fold symmetry of nuclear pores

109
Q

small molecules (5kDa or less) can

A

rapidly diffuse between cytoplasm and nucleoplasm

110
Q

proteins of 20-40,000 Da

A

diffuse more slowly

111
Q

proteins >40kDa cannot enter

A

and RNA/ribosomes cannot exit unless they carry nuclear localisation or export signals

112
Q

22% of human proteins are

A

nuclear and must be imported from cytosol

113
Q

diffusion barrier is caused by

A

unstructured regions of NPC proteins forming tangled network blocking the passive diffusion of large molecules

114
Q

NPC is

A

125 x 10^6and has 30 different nucleoporins`

115
Q

nuclear localisation signals (NLS)

A

rich in lysine and proline and can be in any position of the passenger (cargo) proteins

so long as they are exposed to the surface of the protein

116
Q

importins are the NLS receptors

A

recognised by a family of cytosolic nuclear import receptors each member being responsibsible for a set of cargo molecules

117
Q

nuclear import

A

since transport is through large pores, fully folded proteins and newly formed ribosomal subunits can be transported in and out respectively

the importing binds the NLS and the FG repeats in the FG nucleoporins of the fibrils and channel

118
Q

how does importing know when to let go of its cargo

A

-GTPase switch
-a protein that can either bind GTP or GDP and can hydrolyse GTP to GDP

and assumes a different conformation depending on which nucleotide it binds
-different conformation = different activity

119
Q

Ran-GDP has conformation

A

A and in this state only in cytosol

120
Q

Ran-GTP

A

has conformation B and in this state is only in nucleus

121
Q

Ran-GTP arriving in cytosol is

A

immeditly converted

122
Q

extracellular matrix (ECM)

A

supports cells and their influences
-strength
-elastcity
-turgor

123
Q

Gl;ycosaminoglycan (GAG)

A

chains are repeated units of negatively charged dissacharides that form linear chains
they attract cations(Na+) causing large amounts of water to be sucked into the matrix
they occupy a large volume relative too their mass and creat turgor

124
Q

negatively charged attracts cat ions so

A

osmosis occurs

125
Q

an aggrecan aggregate

A

Is composed of aggrecan bound to hyaluronan

-a proteoglycan with 100+ GAG chains

126
Q

aggrecan molecular weight

A

2.10^8

127
Q

proteoglycans(PGs) that contain GAGs

A

means pore sizes vary
they can bind signalling molecules or can bind to proteases to concentrate them

128
Q

fibroblasts in the ECM

A

what you see mostly is fibrillar collagen
about 25% of protein in body is collagen

129
Q

fibrillar collagen structure

A

N terminal polypeptide (150 aa)
signal peptide(SP) directs the nascent protein
glycine residues (the repetitive structure promotes trimerisation)
-C terminal propeptide(250 aa)

130
Q

modifications in ER

A

1)about 50% of the proline residues are hydroxylated (-OH) by collagen propel-hydroxylases this requires vitamin C
2)specific lysine residues are hydroxylated this also requires vitamin C
3)the c terminal domains are N glycosylated
4)and mostly in the Golgi some of the hydroxylsysl are O glycosylated by addition of galactose or glucose galactose

131
Q

winding production triple stranded helical structure

A

with highly soluble N and C termini

132
Q

function of N and C termini

A

-one other function is to keep trimers apart to stop them forming fibrils
-until late the secretory pathway when the pro peptides are removed forming tropocollagen

133
Q

fibripositors

A

individual tropocollagen are cross linked to form fibrils

134
Q

the type IX collagen is a fibril-associated collagen that can interact with type II

A

this interacts with type 1 collagen fibrez

135
Q

elastin

A

a hydrophobic elastic protein with extensive crosslinks

highly prevelant in ECM of arteries gives tissue their elastcicty

136
Q

cytoskeleton

A

-provides mechanical strength
-drives organelle movement
-severs as an anchor for cell-cell junctions
-drives chromosomes segregation in mitosis and splits the cell in two(cytokinesis)
-enables cell movement and muscle contraction

137
Q

microtubules

A

non covalent heterodimer of alpha and beta subunits
-both subunits bind GTP
-there is polarity a += end and - end
-GTP bound heterodimers bind only at ends growth of microtubule is unidirectional from the + end only
-microtubules are stiff stuctures

138
Q

growth of micro

A

-grows/shrinks only at + end
-they are dynamically unstable

139
Q

actin filaments

A

-non covalent polymers of actin monomers
-each monomer binds ATP (which is replaced by Ap in the filament)
-two filaments twist around each other to form an actin molecule
-there is a polarity a plus and minus end

140
Q

IFs are rope like fibres ,Ade of intermediate filament

A

proteins that are made of alpha helical monomers that associate into coiled coils that then associate in filaments
-IFs form a very large diverse family of non-nucleotide binding proteins including the nuclear laming and epithelial keratins
-intermediate filaments function = they provide mechanical strength they bend but do not break

141
Q

types of junctions

A

**

142
Q

adhering form

A

HOMODIMERS WITH THE EXTRACELLULAR PART OF EACH POLYPEPTIDE FOLDED INTO FIVE CADHERIN REPEATS
THERE ARE CA 2+ BINDING SITES BETWEEN EACH BPAIR OF REPEATS

143
Q

cadherins and homophillic binding

A

-interact weakly so each pair is disassembled

144
Q

Wilson HVP (1907)

A

-on some phenomena of coalescence and regeneration in sponges

-they were disaggregated into single cells by forcing pieces of sponge through a fine sieve

-tried to make a hybrid from two different species by disaggregating cells and mixing them but no hybrids so sponges have HOMOTYPIC recognition

145
Q

adherent junctions summary

A

-the cell to cel interacting membrane proteins are cadherins
-the homophilllic interactions they make are weak, but many acting together makes for strong attachment proteins
-enable cells to be recognised
-junctions connect indirectly to actin cytoskeleton and influence the co-ordinated contractions of cells
-complex interplay between chemical signalling pathways and cell-cell adhesion

146
Q

integrins

A

transmembrane proteins that allow the internal cytoskeleton to grip onto molecules of ECM when necessary

147
Q

integrins

A

transmembrane proteins that allow the internal cytoskeleton to grip onto molecules in ECM when necassary

148
Q

interns are receptors for ECM molecules

A

-a alpha/beta heterodimerthat links an ECM to talin
-talin links to actin other proteins reinforce the linkage
-integrins there are 8 different beta chains and 18 different alpha chains each with distinct ligand binding prop[erties `

149
Q

junctions

A

**

MCO (2 decks)

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