Microscopy and Cell Structure

Chapter 3
What were the primary take
home messages of the last
chapter (2)?
Atoms, elements and molecules comprise living systems

Elements are joined by chemical bonds

Which element is more reactive - C, H or O, and why?

4 major biological molecules

Which ones used for energy

Which are most common carbohydrates on the planet?
What is a molecule?

Molecules <1 nm
 Question of the Day*:
Which organelle is
derived from a microbial
ancestor?
 Microscopes
Microscopes
Most important tool for
studying microorganisms
Use viable light to observe
objects
Magnify images
approximately 1,000x
Electron microscope,
introduced in 1931, can
magnify images in excess
of 100,000x
Scanning probe
microscope, introduced in
1981, can view individual
atoms
 Principles of Light Microscopy
Light Microscopy
Light passes through specimen, then through
series of magnifying lenses
Most common and easiest to use is the bright-
field microscope
Important factors in light microscopy include
Magnification
Resolution
Contrast
 Magnification
Microscope has two magnifying lenses
Called compound microscope
Lens include
Ocular lens and objective lens
Most bright field scopes have four magnifications of
objective lenses, 4x, 10x, 40x and 100x
Lenses combine to enlarge objects
Magnification is equal to the factor of the ocular x
the objective

10x X 100x = 1,000x

Focal Path
 Principles of Light Microscopy
Magnification
Bright field scopes have condenser lens
Has no affect on magnification
Used to focus illumination on specimen
 Resolution
Usefulness of a
microscope depends on
its ability to resolve two
objects that are very
close together

Resolving power is
defined as the minimum
distance existing
between two objects
where those objects still
appear as separate
objects
Resolving power
determines how much
detail can be seen
 Resolution
Resolution depends on the quality of lenses
and wavelength of illuminating light
How much light is released from the lens
Maximum resolving power of most brightfield
microscopes is 0.2 m (1x10-6)
This is sufficient to see most bacterial structures
Too low to see viruses
Principles of Light Microscopy
Resolution
Resolution is enhanced with
lenses of higher magnification
(100x) by the use of immersion oil
Oil reduces light refraction
Light bends as it moves from glass to
air
Oil bridges the gap between the
specimen slide and lens and reduces
refraction
Immersion oil has nearly same
refractive index as glass
 Principles of Light Microscopy
Contrast
Reflects the number of visible shades in a
specimen
Higher contrast achieved for microscopy
through specimen staining
 Principles of Light Microscopy
Examples of light microscopes that
increase contrast
Phase-Contrast Microscope
Interference Microscope
Dark-Field Microscope
Fluorescence Microscope
Confocal Scanning Laser Microscope
 Principles of Light Microscopy
Phase-Contrast
Amplifies differences between refractive indexes of
cells and surrounding medium
Uses set of rings and diaphragms to achieve
resolution
 Principles of Light Microscopy
Interference Scope
This microscope causes
specimen to appear three
dimensional
Depends on differences in
refractive index
2 beams of light pass through
Specimen and then recombine
Most frequently used
interference scope is Nomarski
differential interference contrast
 Principles of Light Microscopy

Dark-Field Microscope
Reverse image
Specimen appears bright
on a dark background
Like a photographic
negative
Achieves image through
a modified condenser;
Light is directed at an angle
 Principles of Light Microscopy
Fluorescence Microscope
Used to observe organisms
that are naturally fluorescent
or are flagged with
fluorescent dye
Fluorescent molecule absorbs
ultraviolet light and emits
visible light
Image fluoresces on dark
background

FISH - fluorescent in situ hybridization

https://www.facebook.com/quartznews/videos/1074441669256201/
A. tubulata with V. Shiloi AK1 probe

By K. Negandhi 2009
 Confocal Microscopy
Confocal Scanning Laser
Microscope
Used to construct three
dimensional image of
thicker structures
Provides detailed sectional
views of internal structures
of an intact organism
Laser sends beam through
sections of organism
Computer constructs 3-D
image from sections

http://www.epa.gov/nerlcwww/images.htm
http://www.microbes.info/resources/General_Microbiology/Imag
es/index.html
http://www.home.duq.edu/~stolz/RIBS/lab/confocal/confo.htm
 Principles of Electron
Microscopy
Electron Microscope
Uses electromagnetic lenses, electrons and
fluorescent screen to produce image
Resolution increased 1,000 fold over
brightfield microscope
To about 0.3 nm (1x10-9)
Magnification increased to 100,000x
Two types of electron microscopes
Transmission
Scanning
http://www.inano.dk/research/co
mpetences-and-
facilities/nanotools/transmission-
and-scanning-electron-
microscopy/
 Transmission Electron
Microscopy
Transmission Electron
Microscope (TEM)
Used to observe fine detail
Directs beam of electrons at
specimen
Electrons pass through or scatter at
surface
Shows dark and light areas
Darker areas more dense
Specimen preparation through
Thin sectioning
Freeze fracturing or freeze etching
Sponges : Choanosome (inside)

Intracellular bacteria

Bacteriocytes
Scanning Electron Microscope (SEM)
Electron Microscopy
Scanning Electron
Microscope (SEM)
Used to observe surface detail
Beam of electrons scan
surface of specimen
Specimen coated with metal
Usually gold
Electrons are released and
reflected into viewing chamber
Some atomic microscopes
capable of seeing single atoms
mcb.arizona.edu

nimr.mrc.ac.uk
Human
Eyelashes

http://www.chemistryexplained.com/images/chfa_02_img0322.jpg
electrodeposited magnetic nanowire array
http://image.mrs.org/images/f07scienceasart/F07FirstPlace-FannyBeron_1.jpg
http://geoinfo.amu.edu.pl/wpk/pe/a/harbbook/c_i/chap01.html
 A cryo-
electron
micrograph
showing a
single layer of
evenly spaced
enzyme
structures
(colorful
"wheels")
interspersed
with gold
nanoparticles
(magenta)

http://www.bnl.gov/bnlweb/pubaf/pr/PR_display.asp?prID=07-73
 Atomic Force microscopy

http://pubs.rsc.org/en/content/articlelanding/2006/ob/b605464a
AFM
 Dyes and Staining
Cells are frequently stained to observe organisms
Stains are made of organic salts

Dyes carry either a (+) or (-) charge on the molecule

Molecule binds to certain cell structures

Dyes divided into basic or acidic based on charge (Opposites

Attract!)
Basic dyes carry positive charge and bond to cell
structures that carry negative charge
Commonly stain the cell
Acidic dyes carry negative charge and are repelled by cell structures that carry
negative charge
Commonly stain the background

Some basic dyes:

Methylene blue,
crystal violet, safranin,
malachite green
 Microscope Techniques
Dyes and Staining
Staining Procedures
Simple stain uses one basic stain to stain the
cell
Allows for increased contrast between cell and
background
All cells stained the same color
No differentiation between cell types
 1. Cryptosporidi
um parvum
oocysts

1. Differential interference contrast (DIC) image of

2. Immunofluorescence detection by antibodies
3. DAPI - DNA staining
http://www.epa.gov/nerlcwww/cpt_seq1.htm
 Dyes and Staining
Differential Stains
Used to distinguish one bacterial group from
another
Uses a series of reagents
Two most common differential stains
Gram stain
Acid-fast stain
 Dyes and Staining
Gram Stain
Most widely used procedure for staining bacteria
Developed over century ago
Dr. Hans Christian Gram
Bacteria separated into two major groups
Gram positive
Stained purple
Gram negative
Stained red or pink
 Dyes and Staining
Gram Stain - (you should know
these basic steps*)
Involves four reagents
Primary stain
Crystal violet
Stains the cell
Mordent
Grams iodine
Holds primary dye onto cell
Decolorizer
Usually alcohol
Removes primary dye from
Gram negative cell
Counter or Secondary stain
Safrinin
Recolors cells that lose stain
through decolorization
 Dyes and Staining
Acid-fast Stain
Used to stain organisms that resist
conventional staining
Used to stain and IDENTIFY members of
genus Mycobacterium
High lipid concentration in cell wall prevents
uptake of dye
Uses heat to facilitate staining
Once stained difficult to decolorize
 Dyes and Staining
Acid-fast Stain
Can be used for presumptive
identification in diagnosis of
clinical specimens
Requires multiple steps
Primary dye
Carbol fuchsin
Colors acid-fast
bacteria red
Decolorizer
Generally acid alcohol
Removes stains from
non acid-fast bacteria
Counter stain
Methylene blue
Colors non acid-fast
bacteria blue
Dyes and Staining - can help ID
specific cellular structures
Special Stains
Capsule stain
Example of negative stain
Allows capsule to stand out around
organism
Endospore stain
Staining enhances endospore
Uses heat to facilitate staining
Flagella stain
Staining increases diameter of
flagella
Makes more visible
 Morphology or shape of
Prokaryotic Cells
Prokaryotes exhibit a
variety of shapes
Most common
Coccus
Spherical
Bacillus
Rod or cylinder
shaped
Cell shape not to
be confused with
Bacillus genus
Morphology of Prokaryotic Cells
Prokaryotes exhibit a
variety of shapes
Other shapes
Coccobacillus
Short round rod
Vibrio
Curved rod
Spirillum
Spiral shaped
Spirochete
Helical shape
Pleomorphic
Bacteria able to vary
shape
Comparison of Eukaryotes to Prokaryotes (see
alsoTable 3.7)

http://learn
somescien
ce.com/
Morphology of Prokaryotic Cells
Prokaryotic cells may form groupings after
cell division
Cells adhere together after cell division for
characteristic arrangements
Arrangement depends on plan of division
Especially in the cocci
Morphology of Prokaryotic Cells
Division along a single plane may result in pairs
or chains of cells
Pairs = diplococci
Example: Neisseria gonorrhoeae
Chains = streptococci
Example: species of Streptococcus

Attention to
Details!
Morphology of Prokaryotic Cells

Division along two or three perpendicular planes

form cubical packets
Example: Sarcina genus
Division along several random planes form
clusters
Example: species of Staphylococcus (see lab chp 2 of
lab manual for details - useful for identification)
Morphology of Prokaryotic Cells
Some bacteria live in groups with
other bacterial cells
They form multicellular
associations
Example: myxobacteria
These organisms form a
swarm of cells
Allows for the release of
enzymes which degrade
organic material
In the absence of water
cells for fruiting bodies
Other organisms for biofilms
Formation allows for
changes in cellular activity
 Cytoplasmic membrane
Defines boundary : in vs out
Serves as a semi permeable barrier
Barrier between cell and external environment
Cytoplasmic membrane
Defines cell boundary
Serves as a semi permeable barrier
Barrier between cell and external environment
Structure is a lipid
bilayer with embedded
proteins
Bilayer consists of two
opposing leaflets
Leaflets composed of
phospholipids
Each contains a
hydrophilic phosphate
head and hydrophobic
fatty acid tail
 Cytoplasmic Membrane
Membrane is embedded
with numerous protein
More that 200 different
proteins
Proteins function as
receptors and transport
gates
Provides mechanism to
sense surroundings
Proteins are not stationary
Constantly changing
position
Called fluid mosaic model
 Cytoplasmic membrane is selectively
permeable
Determines which molecules pass into or out
of cell
Few molecules pass through freely
Molecules pass through membrane via
simple diffusion or transport mechanisms
that may require carrier proteins and
energy
 Simple diffusion
Process by which molecules move freely
across the cytoplasmic membrane
Water, certain gases and small hydrophobic
molecules pass through via simple diffusion
Osmosis
The ability of water to
flow freely across the
cytoplasmic membrane
Water flows to equalize
solute concentrations
inside and outside the
cell
I-Chip method to obtain more uncultivable bacteria
Nichols et al, Appl Env Microbiol (see course content
 Simple diffusion
Inflow of water exerts
osmotic pressure on
membrane
Membrane
rupture is
prevented by
rigid cell wall of
bacteria

http://www.youtube.com/watch?v=sdiJtDRJQEc
Directed movement across the
membrane
Movement of many molecules
directed by transport systems
Transport systems employ highly
selective proteins
Transport proteins (a.k.a permeases
or carriers)
These proteins span
membrane
Single carrier transports
specific type molecule
Most transport proteins are
produced in response to need
Transport systems include
Facilitated diffusion
Active transport
Group translocation
 Cytoplasmic Membrane
Facilitated diffusion
Moves compounds across membrane
exploiting a concentration gradient
Flow from area of greater concentration to area of
lesser concentration
Molecules are transported until equilibrium is reached
No energy is required for facilitated diffusion
Example: movement of glycerol into the cell
 Cytoplasmic Membrane
Active transport
Moves compounds against a concentration
gradient
Requires an expenditure of energy
Two primary mechanisms
Proton motive force
ATP Binding Cassette (ABC) system
 Proton motive force
Transporters allow protons
into cell
Protons either bring in or
expel other substances
Example: efflux pumps
used in antimicrobial
resistance
ATP Binding Cassette
system (ABC transport)
Use binding proteins to
scavenge and deliver
molecules to transport
complex
Example: maltose transport
 ATP Binding Cassette system (ABC transport)
Use binding proteins to scavenge and deliver molecules to
transport complex
Example: maltose transport

http://faculty.ccbcmd.edu/courses/bio141/lecguide/unit1/prostruct/ABC_flash.html
 Regulated Flow of Molecules
has impact on cellular activities
Membrane also the site of
energy production in
mitochondria and some
bacteria
Energy (ATP) produced
through series of
embedded proteins
Electron transport chain
Proteins are used in the
formation of proton motive
PMF force (chap 6)
 Cytoplasmic Membrane
Group transport
Transport mechanism that
chemically alters molecule
during passage
Uptake of molecule does not
alter concentration gradient
Phosphotransferase system
example of group transport
mechanism
Phosphorylates sugar
molecule during transport
Phosphorylation changes
molecule and therefore
does not change sugar
balance across the
membrane
 Bacterial cell wall
Rigid structure
Surrounds cytoplasmic membrane
Determines shape of bacteria
Holds cell together
Prevents cell from bursting
Unique chemical structure
Distinguishes Gram positive
from Gram-negative
 Cell Wall
Rigidity of cell wall is due to
peptidoglycan (PTG)
Compound found only in bacteria
Basic structure of peptidoglycan
Alternating series of two subunits
N-acetylglucosamine (NAG)
N-acetylmuramic acid (NAM)
Joined subunits form glycan chain
Glycan chains held together by string
of four amino acids
Tetrapeptide chain
Gram+ Cell Wall
Gram positive cell wall
Relatively thick layer of
PTG
As many as 30
Regardless of
thickness, PTG is
permeable to
numerous substances
Teichoic acid
component of PTG
Gives cell negative
charge
Gram+ Cell Wall
 Gram-negative Cell Wall

More complex than Gram+

Only contains thin layer of PTG
PTG sandwiched between
outer membrane and
cytoplasmic membrane
Region between outer
membrane and cytoplasmic
membrane is called periplasm
Most secreted proteins
contained here
Proteins of ABC transport
system located here
 Gram-neg Cell Wall
Outer membrane
Constructed of partial lipid bilayer
Much like cytoplasmic membrane but outer leaflet made of
lipopolysaccharides not phospholipids
Outer membrane also called the lipopolysaccharide layer or
LPS layer

LPS serves as barrier

to a large number of
molecules
Replaces one
phospholipid layer
Small molecules or
ions pass through
channels called porins
 Medically important LPS portions:
O-specific polysaccharide side chain
Directed away from membrane
Opposite location of Lipid A
Used to identify certain species or strains
Pathogenic E. coli O157:H7 refers to specific O-side chain

Lipid A
Portion that anchors LPS
molecule in lipid bilayer
Plays role in recognition
of infection
Molecule present with
Gram negative infection
of bloodstream
 Cell Wall
PTG as a target for antibiotics
Many antimicrobials interfere with the
synthesis of PTG
Examples include
Penicillin
Lysozyme
 Cell Wall
Penicillin
Binds proteins involved in cell wall synthesis
Prevents cross-linking of glycan chains by
tetrapeptides
More effective against Gram positive
bacterium
Due to increased concentration of PTG
Penicillin derivatives produced to protect against
Gram negatives
 Lysozymes
Produced in many body fluids including tears
and saliva
Breaks bond linking NAG and NAM
Destroys structural integrity of cell wall
Enzyme often used in laboratory to remove
PTG layer from bacteria
Produces protoplast in G+ bacteria
Produces spheroplast in G- bacteria
 Cell Wall
Differences in cell wall account for
differences in staining characteristics
Gram-positive bacterium retain crystal violet-
iodine complex of Gram stain
Gram-negative bacterium lose crystal violet-
iodine complex
 Cell Wall
Some bacterium naturally lack cell wall
Mycoplasma
Bacterium causes mild pneumonia; small genome
Have no cell wall
Antimicrobial directed towards cell wall ineffective
Sterols in membrane account for strength of membrane
Bacteria in Domain Archaea
Have a wide variety of cell wall types
None contain peptidoglycan but rather
pseudopeptidoglycan
 Question of the Day*:

Do microbes sleep?
 Layers External to Cell Wall
Capsules and Slime Layer
General function
Protection
Protects bacteria from host defenses
Attachment
Enables bacteria to adhere to specific
surfaces
Capsule is a distinct gelatinous layer
Slime layer is irregular diffuse layer
Chemical composition of capsules and
slime layers varies depending on
bacterial species
Most are made of polysaccharide
Referred to as glycocalyx
Glyco = sugar calyx = shell
 Bacterial Spores
Endospores
Dormant cell types
Produced through sporulation
Theoretically remain dormant for
100 years
Resistant to damaging conditions
Heat, desiccation, chemicals and
UV light
Vegetative cell produced through
germination
Germination occurs after
exposure to heat or chemicals
Germination not a source of Common bacteria genus that
reproduction produce endospores include
Clostridium and Bacillus
 Internal Structures
Endospore formation
Complex, ordered sequence
Bacteria sense starvation and begin
sporulation
Growth stops
DNA duplicated
Cell splits
Cell splits unevenly
Larger component engulfs small component,
produces forespore within mother cell
Forespore enclosed by two membranes
Forespore becomes core
PTG between membranes forms core wall
and cortex
Mother cell proteins produce spore coat
Mother cell degrades and releases
endospore
 Flagella and Pili
Some bacteria have protein appendages
Not essential for life
Aid in survival in certain environments
They include
Flagella
Pili
 Flagella and Pili
Flagella
Long protein structure
Responsible for motility
Use propeller like
movements to push
bacteria
Can rotate more than
100,00 revolutions/minute
82 mile/hour
Some important in bacterial
pathogenesis
H. pylori penetration
through mucous coat
 Flagella and Pili
Flagella structure has
three basic parts
Filament
Extends to exterior
Made of proteins called
flagellin
Hook
Connects filament to cell
Basal body
Anchors flagellum into cell
wall

http://www.youtube.com/watch?v=q1iCjKWzeEE&feature=related
 Flagella and Pili
Bacteria use flagella for
motility
Motile through sensing
chemicals, or light
Chemotaxis

If chemical compound is
nutrient
Acts as attractant
If compound is toxic
Acts as repellent
Flagella rotation
responsible for run and
tumble movement of
bacteria
Flagella and Pili
Pili
Considerably shorter and
thinner than flagella
Similar in structure
Protein subunits
Function
Attachment
These pili called fimbre
Movement
Conjugation
Mechanism of DNA
transfer
Flagella in DIC

Cells of E. coli were examined by video-enhanced differential-interference-

contrast microscopy. The depth of field is shallow, and all the cells are near the
bottom of the preparation. Some cells are shown de-energized, exhibiting
filaments with a variety of wave forms: normal, coiled, semi-coiled, curly 1, and
curly 2. See Block, S.M., Fahrner, K.A. and Berg, H.C. "Visualization of
bacterial flagella by video-enhanced light microscopy." J. Bacteriol. 173, 933-
936 (1991).

http://www.rowland.harvard.edu/labs/bacteria/movies_misc.html
Swimming Rhodobacter spheroides
Introduction
This organism has one flagellar filament that emerges from the
side of the cell body. When the filament spins (usually
clockwise), it is helical and the cell runs. When the filament
stops, it coils up and the cell stops. The coiling tends to reorient
the cell body.
 Twitching in Pseudomonis aeruginosa
Introduction

Type IV pili are thin (6 nm dia.) filaments that are essential for twitching
motility in Pseudomonas aeruginosa and social gliding in Myxococcus
xanthus. How do Type IV pili generate cell movement? The first clues
came from electron microscopy studies of bacteriophage infection of a
variety of Gram-negative bacteria. After the initial interaction between the
phage and the pilus, it appeared that retraction pulled the phage to the cell
surface where productive infection could occur.

http://www.rowland.harvard.edu/labs/bacteria/movies/paeru_f_
move_1.mov
 Internal Structures
Bacterial cells have variety of internal structures
Some structures are essential for life
Chromosome
Ribosome
Others are optional and can confer selective
advantage
Plasmid
Storage granules
Endospores
 Internal Structures
Chromosome
Resides in cytoplasm
In nucleoid space
Typically single chromosome
Circular double-stranded molecule
Contains all genetic information
Plasmid
Circular DNA molecule
Generally 0.1% to 10% size of
chromosome
Extrachromosomal
Independently replicating
Encode characteristic
Potentially enhances survival
Antimicrobial resistance
 Ribosome
Involved in
protein
synthesis
Composed of large and small
subunits
Units made of protein and
ribosomal RNA
Prokaryotic ribosomal subunits
Large = 50S
Small = 30S
Total = 70S
Larger than eukaryotic
ribosomes
40S, 60S, 80S
Difference often used as
target for antimicrobials
Small
Subunit
rRNA

rRNA of Thermus
thermophilus and the
archaeon Haloarcula
marismort
3 Domains
Of Life
(based on
16S rRNA
Gene
Sequences)
 Internal Structures
Storage granules
Accumulation of polymers
Synthesized from excess
nutrient
Example = glycogen
Excess glucose in cell is
stored in glycogen
granules
Gas vesicles
Small protein compartments
Provides buoyancy to cell
Regulating vesicles allows
organisms to reach ideal
position in environment
http://scienceblogs.com/afarensis/upload/2006/01/magnetosomes_xl.jpg
Eukaryotes
Comparison of Eukaryotes to Prokaryotes (see
alsoTable 3.7)

http://learn
somescien
ce.com/
 Eukaryotic Plasma Membrane
Similar in chemical structure and function of cytoplasmic
membrane of prokaryote
Phospholipid bilayer embedded with proteins
Proteins in bilayer perform specific functions
Transport
Maintain cell integrity
Attachment of proteins to internal structures
Receptors for cell signaling
Proteins in outer layer
Receptors typically glycoproteins
Membrane contains sterols for strength
Animal cells contain cholesterol
Fungal cells contain ergosterol
Difference in sterols target for antifungal medications
 Eukaryotic Plasma Membrane
Transport across eukaryotic membrane
Some molecules pass through membrane via
transport proteins
Others taken in through endocytosis and
exocytosis
 Eukaryotic Plasma Membrane
Transport proteins
Function as carriers or channels
Channels create pores in membrane
Channels are gated
Open or closed depending on environmental conditions
Concentration gradient
Carriers analogous to prokaryotic membrane
proteins
Mediate facilitated diffusion and active transport
 Endocytosis
Process by which eukaryotic cells bring in material from
surrounding environment
Pinocytosis most common type in animal cell
Pinch off small portions of own membrane along with attached
material

Internalize vesicle and contents

Vesicle called endosome
 Eukaryotic Plasma Membrane
Endocytosis
Phagocytosis
Specific type of endocytosis
Important in body defenses
Phagocyte sends out pseudopods to surround microbes
Phagocyte brings microbe into vacuole
Vacuole = phagosome
Phagosome fuses with a sack of enzymes and toxins
Sack = lysosome
Fusion of phagosome and lysosome creates phagolysosome
Microbe dies in phagolysosome
Phagosome breaks down microbial material
 Eukaryotic Plasma Membrane
Exocytosis
Reverse of endocytosis
Vesicles inside cell fuse with plasma
membrane
Releases contents into external environment
 Protein Structures of
Eukaryotic Cell
Eukaryotic cells have unique structures
that distinguish them from prokaryotic
Cytoskeleton
Flagella
Cilia
80s ribosome
 Protein Structures of
Eukaryotic Cell
Cytoskeleton
Threadlike proteins
Reconstructs to adapt
to cells changing
needs
Composed of three
elements
Microtubules
Actin filaments
Intermediate fibers
 Protein Structures of
Eukaryotic Cell

Microtubules
Thickest of cytoskeleton structures
Long hollow cylinders
Protein subunits called tubulin
Form mitotic spindles
Main structures in cilia and flagella
 Ancient genes and proteins

Like tubulin - a cytoskeletal protein

Organisms Diversity & EvolutionVolume 3,
Issue 3, 2003, Pages 185-194

http://www.cellbio.duke.edu/faculty/Ericksoan/nucleationgTuRC.JPG
Complete primary sequence of tubulin protein
Comparison of protozoan and human tubulin protein
sequences show 82% match
 Evolution:
Nothing in biology makes sense
Except in the light of Evolution
-T. Dobzhansky
the organism is a survival
machine built by short-lived
confederations of long-lived
genes.

- Richard Dawkins
 DISCODERMOLIDE

Untreated cancer cells

H
O O

OH OH O
O
OH OH Cancer cells treated with
NH2
Licensed by HBOI to Novartis discodermolide.
for cancer drug development; Micrographs courtesy of
Structure by S Gunasekera Dr. Ross Longley, HBOI
 Protein Structures of
Eukaryotic Cell
Actin filaments
Composed of actin polymer
Enable cell cytoplasm to move
Assembles and disassembles causing motion
Pseudopod formation
 Protein Structures of
Eukaryotic Cell

Intermediate fibers
Function to strengthen cell
Enable cells to resist physical stress
 Protein Structures of
Eukaryotic Cell
Flagella
Flexible structure
Function in motility
9+2 arrangement
9 pairs of microtubules
surrounded by 2 individual
Cilia
Shorter than flagella
Often cover cell
Can move cell or propel
surroundings along stationary
cell
Intracellular Bacteria
 Membrane-bound Organelles
of Eukaryotes
Eukaryotes have numerous organelles
that set them apart from prokaryotic cells
Nucleus
Mitochondria and chloroplast
Endoplasmic reticulum
Golgi apparatus
Lysosome and peroxisomes
 Membrane-bound Organelles
of Eukaryotes
Nucleus
Distinguishing feature of
eukaryotic cell
Contains DNA
Area of DNA replication
Mitosis = asexual
Meiosis = sexual
Mitochondria
Site of energy production
Surrounded by membrane
bilayer
Inner and outer membrane
Outer membrane
invaginations called cristae
Matrix formed from inner
membrane
Contains its own DNA,
genome
 Felis catus mtDNA
genome - 17009 bp

Lopez et al, Genomics 33:229-246

 ENDOSYMBIOSIS? : Cat
mitochondrial transposition and
amplification in the nucleus

J Lopez et al (1994) Numt, a recent transfer

and tandem amplification of mitochondrial
DNA in the nuclear genome of the
domestic cat. J. Mol. Evol. 39:171-190.
Membrane-bound Organelles
of Eukaryotes
Chloroplast
Found only in plant and algae
Site of photosynthesis
Surrounded by two
membranes
Endoplasmic reticulum
Divided into rough and smooth
Rough ER
embedded with ribosomes
Site of protein synthesis
Smooth ER
Lipid synthesis and degradation
Calcium storage
 Membrane-bound Organelles
of Eukaryotes
Golgi apparatus
Consists of a series of
membrane bound flattened
sacs
Modifies macromolecules
produced in endoplasmic
reticulum
Lysosomes & Peroxisomes
Lysosomes contain
degradative enzymes
Proteases and nucleases
Peroxisomes
Organelles in which oxygen
is used to oxidize
substances
Breaking down lipids
detoxification
AFM
Microscopic Scales

Molecules <1 nm