1/20/20

HANDS-0N SUMMER-BREAK RESEARCH

DOBBIE SMITH PRIZES
University of Glasgow – Institute of Molecular, Cellular and Systems Biology
and the Plant Science Research Group

£1000 prize bursary

and 4-8 weeks’ summer research experience

Consult an approved member of the MCSB staff, agree a project, then …

submit a letter of application to the Undergraduate School.

Open to all first-year students in the Natural Sciences and allied studies
at the University of Glasgow.

Details available on the MOODLE site

or from Prof. M. Blatt (michael.blatt@glasgow.ac.uk)
The deadline for submissions is 12:00 noon Friday, 20.March 2020

Membrane structure and function

What are polymers? Membranes?

Why are membranes important? Where are

they found?
What are organelles and what are their roles?

How are organelles formed?

What is an electrochemical gradient?

Biology, Campbell&Reese 8th ed., pp. 74-77, 94-135

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What are polymers?

Polymers are molecules built of repeated subunits

Biological polymers are generally formed by dehydration

(water removal)

Common polymers – DNA, RNA, proteins, cellulose, keratin, lipids

What are membranes?

Membranes are phospholipid polymers of fatty acids, glycerol,
phosphate and a terminal amine or alcohol group

Glycerol ester linkage Fatty acid

Phosphatidyl choline comprises

choline, a phosphate linkage to
glycerol which is esterified with
two fatty acids

Hydrophillic head group

Hydrophobic tail groups

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What are membranes?

Membrane phospholipids are amphipathic molecules – in water
they spontaneously form monolayers and bilayers

[Simple detergents are salts of fatty acids (lye soap)

which retain this amphipathicity and are able to
interact and dissolve other organic compounds]

Why are membranes important?

Cellular membranes function …
* to compartmentalize metabolic activities
* to separate/protect cellular components
* to provide a ‘scaffold’ for signalling
* as a medium for cellular energy generation
Compartmentalization, protection and energy generation
arises from the immiscibility of water and membranes

Sucrose diffusion across

membrane bilayers is
1,000,000-fold slower
(10-6 less) still

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What do diffusion rates mean?

How long does it take sucrose to diffuse a distance x?

So the physical properties (diffusion) of compounds in water

limits membrane dimensions and cell size!

Membranes are a barrier to diffusion

Diffusion across semi-permeable membranes
can generate a small imbalance in ion (charge) concentrations

Here the negative charges are meant

to represent proteins (usually carry an
excess of negative charge)

A membrane voltage (electrical charging) occurs when

(i) immobile charges are not balanced
(ii) charge (K+, Cl-, Na+, etc.) transport is not fully balanced

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Biological membranes are often charged

Diffusion of ions (charges) is driven by both chemical and
electrical gradients

The Nernst Equation describes

the equilibrium between
chemical and electrical forces
on a charged molecule (ion)

Ex = -(RT/zF).ln([X]in/[X]out)

Ex = -60mV.log10([X]in/[X]out)

You will learn more of membrane voltage and ion movement tomorrow

Biological membranes are often charged

The voltage across cell membranes can be measured
using microelectrodes implanted in cells

The voltage across other

membranes (eg. mitochondria,
lysosomes) can be measured
with voltage-sensitive dyes

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How big are cells?

Small enough for mean diffusion times

<0.1 s internally and close proximity
of the cytosol to the outside

(N.B. plant cells achieve larger size

with vacuolar space and cyclosis)

Fantastic video:
https://www.youtube.com/watch?v=0fKBhvDjuy0

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Where are membranes found?

Delimiting all organelles

in eukaryotic cells

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Where are membranes found?

mitochondria
chloroplast vacuole
nucleus

Golgi endoplasmic
reticulum

Electron microscopy gives a static picture of the cell

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How are membranes maintained?

Membrane and cargo

delivery/recovery
by endocytosis and
exocytosis

Vesicle traffic through

the Golgi

Membrane lipid synthesis

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SNAREs are key elements in vesicle fusion

Soluble N-ethylmaleimide sensitive factor Attachment protein REceptor

SNAREs ‘anchor’ to target and vesicle

SNAREs (super-)coil together

Coiling pulls membrane faces together …

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Priming the fusion cycle

Sec1/Munc18 proteins prevent non-specific SNARE interactions and
facilitate fusion
NSF (ATPase) and SNAP ‘recharge’ used SNAREs (requires ATP)
Other proteins act in specialised functions … eg. in tethering and
coupling fusion to Ca2+

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Fluorescence – a new window on the cell

Jablonski diagram

How does it work?

Light absorbed by a pigment
‘excites’ electrons

Energy is released as light

as the electron relaxes
back to the ground state
(lowest empty electron
shell)

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Green Fluorescent Protein (GFP)

Aquoria victoria (jellyfish) includes genes

for intrinsically-fluorescent proteins

Aequoria victoria

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GFP expresses in plants and animals

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Endoplasmic reticulum – a network of sausages?

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Endoplasmic reticulum is highly mobile

Tobacco epidermal cell

(HDEL-GFP labelled ER)

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Endoplasmic reticulum is highly mobile

Tobacco epidermal cell

(calnexin-paGFP labelled ER)

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SYP121 is a trafficking protein

GFP normally secreted in secGFP-
transfected leaves (here stained
with propidium iodide)

GFP accumulates in ER and Golgi

when co-transfected in leaves
with SYP121ΔC
Plasma membrane

Syp121
Exocytosis

Golgi

Endocytosis Anterograde

Endoplasmic reticulum

Retrograde

Vacuole Nucleus

Cell wall

Geelen, et al. (2002) Plant Cell 14, 387

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Endoplasmic reticulum space is interconnected

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Golgi – a stack of sacks?

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Is the Golgi and interconnected network

in animal cells?

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Are plant Golgi like animal Golgi?

HDEL-GFP marker for ER

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How autonomous are chloroplasts?

Chloroplasts and mitochondria – endosymbiont progenitors

suggested by double membranes

Chlroplasts and mitochondria undergo

independent division/replication

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Chloroplasts communicate within cells

Stromules (Maureen Hanson)

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How big are cells?

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Membrane structure and function

Membranes are phospholipid polymers which are amphipathic

and spontaneously assemble mono/bilayers in water
Membranes serve a number of roles in cells (you’ll hear
more about this tomorrow) as physical barriers and
structures
Organelles are membane-delimited compartments in
eukaryotic cells
Cells and organelles are highly dynamic

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