Fimbriae

Gram-negative bacteria- short, fine, hair-like

appendages
sometimes called attachment pili
Thinner than flagella and not involved in motility
Cell may be covered with up to 1,000 fimbriae
Small size; visible in electron microscope
Slender tubes; composed of helically arranged
protein subunits and are about 3 to 10 nm in
diameter and up to several micrometers long
Some types of fimbriae attach bacteria to solid
surfaces such as rocks in streams and host tissues
 Pili
Appendages made up of pilin proteins
fragile and constantly replaced
Sex pili: 1 10/cell
Differ from fimbriae:
Pili are larger than fimbriae (around 9 to 10 nm in
diameter).
Genetically determined by sex factors or conjugative
plasmids
Required for bacterial mating
Some bacterial viruses attach specifically to receptors
on sex pili at start of their reproductive cycle
 Pili
Type IV pili: assist cells in adhesion
Twitching motility: unusual form of cell motility;
gliding motility, movement along a solid surface
6 nm in diameter; present at poles of rod-shaped
cells
Key colonization factors- certain human
pathogens, eg. Vibrio cholerae and Neisseria
gonorrhoeae
Assists organism to locate specific sites for
attachment to initiate disease process
 Flagella
Thread-like locomotor appendages; extending outward
from plasma membrane and cell wall
Flagella are thin- cannot be observed directly with a
bright-field microscope, must be stained with special
techniques designed to increase their thickness

Monotrichous Amphitrichous
Single polar flagellum Single flagellum on both sides
Example: Vibrio cholerae Example: Alcaligenes faecalis
Lophotrichous Peritrichous
Tufts of flagella at one or both Numerous flagella all over the
sides bacterial body
Example: Spirillum Example: Salmonella typhi, E.
coli
Examples of non-motile bacteria: Most cocci, Shigella, Klebsiella
Flagella
 Flagellar Ultrastructure/Parts of Flagella
Each flagellum consists of three distinct parts- Filament, Hook and
Basal Body
Filament: external to the cell
Hook: short, curved segment; links the filament to its basal body
Basal Body: attached to cytoplasmic membrane by ring-like
structures; embedded in cell
 Flagellar Ultrastructure/Parts of Flagella
Filament:
hollow, rigid cylinder; single protein called
flagellin
molecular weight: 30,000 to 60,000
Filament ends with a capping protein
Some bacteria sheaths surrounding their flagella
Vibrio cholerae has a lipopolysaccharide sheath
Hook: made of different protein subunits
 Flagellar Ultrastructure/Parts of Flagella
Basal body: most complex part
Most Gram ve bacteria: 4 (L, P, M, S) rings in
basal body connected to a central rod
Outer L and P rings associate with LPS and
peptidoglycan layers, respectively
Inner M ring contacts the plasma membrane
Gram positive bacteria: only two basal body rings,
an inner ring connected to the plasma membrane
and an outer one probably attached to
peptidoglycan
 Flagellar Ultrastructure/Parts of Flagella
---
 Flagellar Synthesis
20 to 30 genes are required
In Escherichia coli and Salmonella enterica, over
50 genes are linked to motility
These genes have several functions:
Encoding structural proteins of flagellum and
motor apparatus
Export of flagellar proteins through cytoplasmic
membrane to outside cell
Control of flagellar construction or function
Regulation of many biochemical events taking
place during synthesis of new flagella
 Flagellar Synthesis
Flagellin subunits: transported through the
filaments hollow internal core
Reach tip- subunits spontaneously
aggregate under direction of a special
filament cap so that filament grows at its tip
rather than at base
Excellent example of self-assembly
 Flagellar Synthesis
M ring synthesized first- inserted into CM
Other anchoring proteins - synthesized along
with hook before filament forms
Flagellin molecules synthesized in cytoplasm
pass up- 3-nm channel inside filament and
add on at the terminus to form the mature
flagellum
At the end of growing flagellum a protein
cap exists
 Flagellar Synthesis

Flagellin subunits travel through the flagellar core and attach to the growing tip
 Mechanism of Flagellar Movement
Propeller of a boat motor
Tiny rotary motor- two main components:
the rotor and the stator
Rotor: basal body
Stator: consists of Mot proteins; surround
basal body and function to generate
torque
Energy required: proton motive force
About 1000 protons are translocated per
rotation of flagellum
 Mechanism of Flagellar Movement
Proton turbine model: protons flowing
through channels in Mot proteins exert
electrostatic forces on helically arranged
charges on rotor proteins
Attractions bet +ve and ve charges cause
basal body to rotate as protons flow
though Mot proteins
Mechanism of Flagellar Movement

Motor switch
 Other Mechanisms of Bacterial Locomotion
Gliding and motion by axial filament
contraction
Gliding: movement of bacteria along solid
surfaces by an unknown mechanism
Axial filaments or endoflagella: internally
located filaments; Spirochetes
Present towards the middle from both
ends
Located above peptidoglycan cell wall but
below the outer membrane
 Cell Walls of Archaea
Peptidoglycan: absent from the cell walls of
Archaea
No outer membrane
Pseudomurein: a polysaccharide; similar to
peptidoglycan
Composed of alternating repeats of NAG and N-
acetyl-talosa-minuronic acid;
glycosidic bonds between the sugar derivatives
are -1,3 instead of -1,4
Some Archaea lack pseudomurein; contain other
polysaccharides
 Cell Walls of Archaea
Eg. Methanosarcina species - thick
polysaccharide walls composed of polymers of
glucose, glucuronic acid, galactosamine uronic
acid, and acetate
Extremely halophilic Archaea; eg. Halococcus
contain sulfate
-ve charge on sulfates bind high concentration of
sodium present in the habitats of Halococcus
Helps stabilize the cell wall in such strongly polar
environments
 Archaeal Membranes
Lipids of Archaea contain ether bonds between
glycerol and their hydrophobic side chains
Archaeal lipids lack true fatty acid side chains
Side chains - composed of repeating units of
hydrophobic five-carbon hydrocarbon isoprene
Cytoplasmic membrane:
either glycerol diethers (20-C side chains; phytanyl
group)
or diglycerol tetraethers (40-C side chains)
 Archaeal Membranes
In tetraether lipid- Ends of phytanyl side chains that
point inward from each glycerol molecule are
covalently linked
This forms a lipid monolayer instead of a lipid bilayer
membrane
lipid monolayer membranes: Extremely resistant to
heat denaturation
Present in hyperthermophiles (80 C)
Membranes with a mixture of bilayer and monolayer
character are also possible
 Archaeal Membranes

Glycerophosphates
Phytanyl

Membrane protein

Biphytanyl

Membrane structure in Archaea may be bilayer

or monolayer (or a mix of both)
 Effect of antibiotics and Enzymes on cell wall
Cell wall: protect bacteria against osmotic lysis
Peptidoglycan can be destroyed by certain agents
Enzyme lysozyme: cleaves -1,4-glycosidic bonds
between NAG and NAM in peptidoglycan
Weakens cell wall; water enter cell and cause lysis
Lysozyme: destroys preexisting peptidoglycan
Lysozyme found in animal secretions including tears,
saliva, and other body fluids
Functions as major line of defense against bacterial
infection
 Effect of antibiotics and Enzymes on cell wall
Penicillin also targets peptidoglycan
Penicillin: prevents biosynthesis of peptidoglycan
If bacteria- incubated with penicillin or lysozyme in
an isotonic solution, Gram +ve bacteria converted
to protoplasts; Gram ve to spheroplasts
Wall-less non-viable organisms that do not multiply
Protoplasts: str of Gram +ve bacteria without cell
wall
Continue to grow normally when isotonicity is
maintained even though they completely lack a
wall
 Effect of antibiotics and Enzymes on cell wall
Spheroplasts: Gram ve cells with outer membrane
but without cell wall
Protoplasts and spheroplasts are osmotically
sensitive
Transferred to a dilute solution- lyse due to
uncontrolled water influx
Mycoplasmas: group of pathogenic bacteria that
causes several infectious diseases of humans and
other animals
lack a cell wall; Plasma membranes are stronger than
normal
Mycoplasmas tend to be pleomorphic or variable in
shape
Most mycoplasmas have sterols in their PM
 Effect of antibiotics and Enzymes on cell wall
L-forms:
Wall-less cells can grow and divide, they are
called L forms
First reported by Klieneberger Nobel in cultures of
Streptobacillus monoliformis
Lister Institute, where they were discovered
produced more readily with penicillin than with
lysozyme
Some L forms are stable (do not revert back);
unstable (revert back to cell wall containing state
when stimulus is removed)
Some spontaneous (Streptobacillusmonoliformis);
inducible
 Effect of antibiotics and Enzymes on cell wall
Difficult to cultivate
Required medium: right osmotic strength and
low concentration of agar, inactivated serum and
sucrose
L forms resemble mycoplasma in morphology,
type of growth on agar fried-egg colony
Different from mycoplasma
mycoplasma lack cell wall and have sterols in
their membrane
L forms may have reminiscent of cell wall but do
not have sterols in their membrane
Mycoplasma pneumoniae
 Action of Penicillin
Most penicillins: derivatives of 6-aminopenicillanic acid
Differ from one another wrt side chain attached to its
amino group
Mechanism not completely known
Structures resemble terminal D-alanine found on the
peptide side chain of the peptidoglycan subunit
Penicillins inhibit enzyme catalyzing transpeptidation
reaction because of their structural similarity
Block synthesis of a complete, fully cross-linked
peptidoglycan and lead to osmotic lysis
Penicillins act only on growing bacteria that are
synthesizing new peptidoglycan
 Penicillin Action

-NAM-NAG-
-NAM-NAG-
L-Ala D-Ala
L-Ala D-Ala

D-Glu DAP
D-GluNH2 L-lys
DAP D-Glu
L-lys D-GluNH2
D-Ala L-Ala
D-Ala L-Ala
-NAM-NAG- Peptide interbridge
-NAM-NAG-

Penicillins inhibit enzyme catalyzing transpeptidation

 Action of Penicillin
Penicillins also bind to several penicillin-binding
proteins and may destroy bacteria by activating their
own autolytic enzymes (autolysins)
Murein hydrolases: move through holes, disrupt the
peptidoglycan, and lyse the cell
Penicillin kills bacteria even in absence of autolysins or
murein hydrolases
Penicillin may stimulate special proteins called
bacterial holins to form holes or lesions in plasma
membrane
Lead to membrane leakage and death
 Effect of antibiotics on cell wall
Penicillin is the classic example of an inhibitor of cell
wall synthesis
Other examples include: ampicillin, bacitracin,
cephalosporin, methicillin, oxacillin and vancomycin
Vancomycin: Disrupts peptidoglycan cross-linkage
Bacitracin: Disrupts movement of peptidoglycan
precursors (topical use)
Antimycobacterial agents: Disrupt mycolic acid
synthesis (bactericidal)
Characteristics Gram Positive Gram Negative
Can be decolorized to accept counterstain (safranin) and
Gram Reaction Retain crystal violet dye and stain blue or purple
stain pink or red
Cell Wall Cell Wall is 20-30 nm thick. Cell Wall is 8-12 nm thick.
Cell Wall The wall is Smooth. The wall is wavy.
Peptidoglycan Layer Thick (multilayered) Thin (single-layered)
Teichoic Acids Present in many Absent
Periplasmic Space Absent Present
Outer Membrane Absent Present
Porins Absent Occurs in Outer Membrane
Lipopolysaccharide (LPS) Content Virtually None High
Low (acid-fast bacteria have lipids linked to
Lipid and Lipoprotein Content High (because of presence of outer membrane)
peptidoglycan)
Mesosomes Quite Prominent Less Prominent
Flagellar Structure 2 rings in basal body 4 rings in basal body
Toxin Produced Exotoxins Endotoxins or Exotoxins
Resistance to Physical Disruption High Low
Low (requires pretreatment to destabilize outer
Cell Wall Disruption by Lysozyme High
membrane)
Susceptibility to Penicillin and
High Low
Sulfonamide
Susceptibility to Streptomycin,
Low High
Chloramphenicol and Tetracycline

Inhibition by Basic Dyes High Low

Susceptibility to Anionic Detergents High Low

Resistance to Sodium Azide High Low

Resistance to Drying High Low