1: The Entropy Problem

 Entropy can be considered as disorder in a system

 Disorder occurs spontaneously
 Order requires energy
 In our universe for any spontaneous process the
entropy must increase. (2nd Law of
Thermodynamics)
 Increase in disorder causes an increase in entropy
 Life is highly organised
 Requires a lot of energy
 Metabolism must be fast
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 2: Mass Transport
 Themovement of substances
 DIFFUSION
 only solute shows net movement
 no net movement of bulk phase (solvent)
 movement random (down concentration gradient)

 CONVECTION
 both solute & solvent move
 vectorial
 rate depends upon input of energy

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But surface to volume ratio is
not the only problem;
Think nerve cells?

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Goodbye to the Prokaryote Cell

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3: Eukaryote Cell Cytoskeleton
 Three Structural Elements,
 All protein fibres made up of monomer sub-units
 Microfilaments,
 monomer a globular protein~Actin
 Microtubules,
 monomer a globular protein ~Tubulin
 Intermediate filaments,
 monomer a long chain protein ~IFP

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 4: Cytoskeleton Functions
 Some functions could be inferred from the structure
and distribution of cytoskeleton.
 Provides a scaffold or framework
 Maintain arrangement of internal organelles
 Control external shape of cell

 Better understanding of the dynamic nature of some

cytoskeleton components required the development of
new microscopic techniques.
 Particularly using fluorescence

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Integrated view of cytoskeleton in an animal cell

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 ACTIN INTER MEDIATE
FILAMENTS MICROTUBULES FILAMENTS

Visualised using anti cytoskeleton flourescent labelled antibodies

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5: Fluorescence Microscopy
 Specimen irradiated by intense light
 Light absorbed by the chromophore is re-
emitted as light of a longer wavelength
 Fluorescence is very weak
 Irradiating wavelengths are excluded by filters
 So that only re-emitted light is viewed

 Zero background = high contrast

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 6: Stokes Shift
 Why is light emitted in fluorescence of longer
wavelength than the light absorbed on excitation?
 Excited electrons initially occupy minor higher energy
levels in the first excited state
 Energy in these minor levels is rapidly lost as heat and
the electrons decay to the lowest energy level in the
first excited state
 When the electrons return to the ground state the energy
lost in fluorescence is less than initially absorbed
corresponding to a longer wavelength of light.

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Absorption
Emission

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7: Only a Few Molecules can Fluoresce
 Most naturally available molecules do not
show fluorescence.
 Only certain molecules can fluoresce;
 Requires a stable first excited state.
 Possible where charge is distributed over a
large delocalised electron cloud.
 Typical structures of fluorescent molecules will
be polycyclic and heterocyclic.

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Some typical
fluorescent labels

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Green Fluorescent Protein
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 8: Green Fluorescent Protein
 Protein isolated from jellyfish
 Unusual in that it can show fluorescence
 Very useful because it is a protein
 So the GFP gene can be transferred to other organisms
and expressed together with other genes
 Forming fusion proteins which can be used to label
living cells, identifying the functional location of
proteins.

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 The GFP
Fluorophore

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 9: Functions of Actin filaments
 Mainly
associated with inner face of plasma
membrane
 Support plasma membrane
 Can move membrane by interaction with
membrane bound myosin and using ATP.
 Involved in vesicle formation
 Can also move organelles along membranes.
 Involved in cell crawling on surfaces
 Actin polymerisation pushes membrane

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 10: Functions of Intermediate Filaments
 Found only in animal cells
 Form tough ropes that transmit forces across cells
in tissue layers and hold them together.
 IF from one cell form direct junctions
(desmosomes) at the plasma membrane with IF
from neighbouring cells
 A class of IF called nuclear lamins reinforce the
inner face of the nuclear membrane (the nucleus lacks
other components of the cytoskeleton)

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 11: Functions of microtubules
 Originally identified as part of mitotic spindle in
animals formed from centrioles
 Characteristic 9+0 arrangement of MT triplets
 Next associated with cilia which have organisers
called basal bodies
 Same arrangement as centrioles
 Now known to have key functions for cell
transport and movement throughout the cytoplasm

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Centrioles

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 12: Cilia & Flagella
 Motile structures derived from plasma membrane
 Central core of microtubules doublets in 9+2
arrangement, the axoneme.
 Microtubules interact with dynein using ATP to
produce bending movements
 In non-motile cilia the axoneme has a 9+0 doublet
microtubule arrangement and no dynein.

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 13: MT Dynamic Instability
 Most Microtubules
 Show continuous assembly or breakdown
 By adding or losing tubulin sub-units at the tip
 Controlled by cap proteins & centrosome
 Allows forces to be applied to different parts of a cell

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 14: MTR = MicroTubule Railway
 Somemicrotubules are static, they do not
grow and shrink all the time.
 These microtubules serve cell internal transport
 Act as a railway track
 Vesicles or organelles can bind to motor
proteins which can bind to the MTR
 Using energy from ATP, the motor proteins can
move the vesicles along the microtubule

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 KINESIN

+
-
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 15: Endomembranes
 Plasma membrane
 Maintains cell integrity, recognition
 Controls transport into and out of cell
 Nuclear membrane
 Controls transport into and out of nucleus
 Endoplasmic reticulum
 Compartmentalises cell functions
 Golgi Body
 Modification and packaging of molecules
 Lysosomes, Vesicles

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16: There is only one sort of e.r.!
 Rough e.r doesn’t really exist
 Except when ribosomes bind
 So what is the difference between soluble
and bound ribosomes?

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 17: Protein Translocation
 mRNA translated at bound ribosomes can
be directly injected in e.r. lumen or inserted
in e.r. membrane
 Requires peptide signal sequence
 Recognised by SRP that binds to pore
protein in e.r. membrane
 necessary to permit hydrophilic protein to
cross hydrophobic interior of membrane

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Golgi

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 18: Post-translational modification
 Glycosylation in the Golgi
 Sequential addition (and removal) of
monosaccharides to proteins by enzymes in
the different layers of the Golgi
 Proteins moved from one Golgi sac to another
in vesicles along microtubules
 Produces glycoproteins

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Origin of Endomembranes

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 19: Organelles with DNA
 Mitochondria
 Site of oxidative metabolism
 Chloroplasts
 Site of photosynthesis
 Centrosomes & centrioles
 Organise microtubules
 All contain circular DNA
 Mitochondria and Chloroplasts surrounded by a
double membrane

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Mitochondrion

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 20: Origin of Organelles
 The organelles have many features
characteristic of prokaryote cells
 Could they have originated through
bacterial symbiosis?
 If a bacterial cell was engulfed by an
eukaryotic cell…..

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 21: Organelles of gene expression
 Nucleus
 Nucleolus
 Site of ribosome synthesis
 Chromosomes
 DNA complexed with protein
 Ribosomes
 Attached to endoplasmic reticulum as RER
 Sites of protein synthesis

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 Import of soluble nuclear protei

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 Import of membrane bound
nuclear proteins

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Nuclear Pore

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 22: Plant Cells
 Cell wall, rigid cellulose structure
 Support & protection
 Chloroplasts
 Vacuole
 Central storage compartment

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 23: Cell Walls
 Plant cell wall composed mainly of cellulose (ß1-4
glucan) arranged in organised microfibrils of
entwined polymer chains.
 Primary wall also contains Pectins, water soluble
complex polysaccharides
 Primary wall thin and extensible.
 Secondary wall contains less soluble hemicelluloses
e.g. xyloglucans and insoluble polyphenols, lignin.
 Secondary wall thick, rigid and impermeable

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 24: Cell Wall Synthesis
 Multi sub unit rings of cellulose synthase form
“rosettes”on plasma membrane
 Synthesise extending glucan chain using
cytoplasmic glucose.
 Chain extension moves rosettes along the
membrane guided by microtubules
 Chains combine to form microfibrils in layers
building up the interleaved texture of the cell wall

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