Major components of prokaryotic cells

Dr. Thomas Seviour twseviour@ntu.edu.sg

 two types of cells

Members of the Microbial World

prokaryotic cell relatively simple morphology lacks a true membrane-delimited nucleus Bacteria and Archaea Term prokaryote blurred eukaryotic cell morphologically complex has a true membrane-delimited nucleus Complex cytoskeleton protozoa, algae, fungi, plants and animals

Prokaryotes
Now clear that PROKARYOTE CELLS
are possessed by two different phylogenetic groups

THE BACTERIA THE ARCHAEA

These differ from each other as
profoundly as eukaryotic cells differ from prokaryotic cells

Term prokaryote becoming blurred

Cell Organization Bacteria and Archaea

Common Features
Cell envelope 3 layers Cytoplasm External structures

Bacterial cell morphology

Bacterial Cell Envelope

Plasma membrane Cell wall Layers outside the cell wall

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Plasma membrane
Absolute requirement for all living organisms separation of cell from its environment selectively permeable barrier
some molecules are allowed to pass into or out of the cell transport systems aid in movement of molecules

location of crucial metabolic processes detection of and response to chemicals in

surroundings with the aid of special receptor molecules in the membrane

Fluid Mosaic Model of Membrane Structure

lipid bilayers with floating
proteins amphipathic lipids polar ends (hydrophilic interact with water) non-polar tails (hydrophobic The fluid mosaic of bacterial insoluble in water) membranes membrane proteins
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Membrane Proteins
peripheral
loosely connected to
membrane easily removed

integral
amphipathic
embedded within membrane carry out important functions may exist as microdomains

The fluid mosaic of bacterial membranes

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Asymmetric Membrane Lipids

Phospholipids, such as phosphatidylethanolamine

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Bacterial Lipids
saturation levels of membrane lipids reflect the
maintain fluidity during growth

environmental conditions such as temperature e.g. at low temperatures, more unsaturated lipids to

bacterial membranes lack sterols but do contain

sterol-like molecules, hopanoids stabilize membrane found in petroleum

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Hopanoids
Sterol like (e.g. cholesterol) Natural pentacyclic compounds Hydrophobic tail, hydrophilic head Natural Membrane Insertion
Molecules (MIM) and rigidity

Increase plasma membrane strength Adjust membrane permeability Adaptation to extreme environmental
conditions

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Antimicrobial peptides
Act by inserting themselves in the plasma
membrane of cells

Destabilizing MIM Potent, broad spectrum antibiotics Part of the innate immune response The amino acid composition, charge and size of
some AMPs allows them to attach to and insert themselves into membrane bilayers, thus killing bacteria by membrane disruption

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Different models for membrane lysis by antimicrobial peptides

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Synthetic MIMs
Biocides, e.g phenylene
ethynylene polyelectrolyte oligomers (OPEs) lipid bilayers

Size and charge mimic Insertion into membrane

causes structural damage that allows leakage of water through membrane

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Synthetic MIMs
Transmembrane electron transfer molecules (TETMs) E.g. Polyvinylene stilbene Designed to increase electron transfer across
membrane

Thus enhance performance of microbial fuel cells or

bioelectrochemical systems

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TETMs

But TETMs also perturb the membrane and are biocidal

Ultimate conformation of the POPE/ POPG bilayer predicted by the molecular dynamics simulation after 200 ns following the intercalation of one and four molecules respectively of DSSN+ (A, B), DSBN+ (C, D), and 4FDSBN+ (E, F) at 300 K. A, C and E represent the low concentrations of TETMs, and B, D and F represent high concentrations. The TETMs are shown in blue, the phosphate head group in yellow, the water molecules in redwhite, and POPE/POPG acyl chains as green lines

Thus need to understand membrane

perturbation for intelligent design of enhancement MIMs

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Cell walls of Bacteria

peptidoglycan (murein)
rigid structure that lies just outside the plasma
membrane

Bacteria are divided into two major groups based

on the response to Gram-stain procedure. gram-positive bacteria stain purple; thick peptidoglycan gram-negative bacteria stain pink; thin peptidoglycan and outer membrane staining reaction due to cell wall structure

Without a cell wall = protoplast

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Cell Wall Functions

maintains shape of the bacterium almost all bacteria have one helps protect cell from osmotic lysis helps protect from toxic materials may contribute to pathogenicity

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Gram positive and Gram negative cell wall organization differs

Both possess a PEPTIDOGLYCAN or MUREIN
layer

In GRAM VES we see an OUTER

LIPOPOLYSACCHARIDE MEMBRANE containing LIPID A

In GRAM+VES we see TEICHOIC ACIDS

Peptidoglycan Structure
important component of both grampositive and gram-negative bacteria subunits

polysaccharide formed from peptidoglycan two (1-4)- -linked alternating sugars

form backbone

N-acetylglucosamine (NAG) N-acetylmuramic acid (NAM) In some bacteria no acetyl group or glycolyl substitution

Peptidoglycan subunits and linkers

Attached to NAM are short peptide chains, usually 4
amino acid residues

Serve to link together the glycan chains in several ways Amino acids there are often unusual Include D-isomers and an amino acid found nowhere
else in biological world

form backbone N-acetylglucosamine (NAG) N-acetylmuramic acid (NAM) In some bacteria no acetyl group or glycolyl substitution
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Peptidoglycan subunit

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Strands Are Crosslinked

peptidoglycan strands have a helical
shape

Amino acids serve to link together the

glycan chains in several ways

Together called GLYCAN

TETRAPEPTIDE = BUILDING BLOCK OF PEPTIDOGLYCANS

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Helical structure enables 360 crosslinking

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Linkage of glycan chains

Varied and involves the
tetrapeptides of adjacent chains involves a direct link between 3rd amino acid residue of one chain (dibasic DAP or L-lysine) and 4th amino acid of other chain (D-Alanine) eg E.coli interbridge of several amino acid residues linkage (i.e. CO-NH)

In many Gram ve bacteria

In many Gram +ves involves an Peptide bonds involved in

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GENERAL COMMENTS
PEPTIDOGLYCAN found ONLY in BACTERIA Diaminopimelic acid (DAP) and muramic
acid unique to BACTERIA

DAP common in Gram ve bacteria, but

replaced in many Gram+ve bacteria by LLysine proteins

D-isomers of amino acids not found in

 composed

Gram-Positive Cell Walls

primarily of peptidoglycan amounts of teichoic acids

also contain large

 Found in nearly all GRAM +ve bacteria May help maintain structure, attachment,
protect cell from harmful substances

Teichoic acids

Polyphosphate sugar alcohols Chemically diverse, with glycerol, ribitol,

mannitol teichoic acids now known

Most bacteria have >1 kind in cell wall Some glycerol teichoic acids bound to
cytoplasmic membrane lipids (i.e. LIPOTEICHOIC ACIDS)

Teichoic acid structure

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Gram-Positive cells

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Periplasmic Space of Gram + Bacteria

lies between plasma membrane and cell
wall and is smaller than that of gramnegative bacteria

periplasm has relatively few proteins Peptidoglycan is ~5-10% of cell wall weight enzymes secreted by gram-positive
bacteria are called exoenzymes
aid in degradation of large nutrients
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Gram-Negative Cell Walls

more complex than gram positive consist of a thin layer of peptidoglycan
surrounded by an outer membrane

outer membrane composed of lipids, no teichoic acids periplasmic space differs from G+

lipoproteins, and lipopolysaccharide (LPS)

may constitute 2040% of cell volume many enzymes present in periplasm hydrolytic enzymes, transport proteins and other
proteins
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Gram-Negative outer layers

outer membrane lies
outside the thin peptidoglycan layer

Brauns lipoproteins

connect outer membrane to peptidoglycan

Adhesion sites direct contact between

plasma membrane and outer membrane substances may move directly into cell through adhesion sites 42

 Lipopolysaccharides (LPS) in outer half of

Outer membrane

outer membrane (phospholipids other) i.e. asymmetric

Lipopolysaccharides consist of three parts

lipid A core polysaccharide with ketodeoxyoctanoic acid
(KDO) and range of sugars unusual dideoxy sugars

O side chain (O somatic antigen) contains

Buried in outer membrane

Lipolysaccharide
Lipid A Lipid A
buried in membrane and core are straight chain bent at an angle

O-side

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Other characteristics of outer membrane

more permeable than plasma membrane due
porin proteins form channels through which small
molecules (600-700 daltons) can pass

to presence of porin proteins and transporter proteins

-barrel structure

Importance of LPS
contributes to negative charge on cell
surface

helps stabilize outer membrane structure may contribute to attachment to surfaces

and biofilm formation

creates a permeability barrier protection from host defenses (O antigen) can act as an endotoxin (lipid A)
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Osmotic Protection (cell wall)

hypotonic environments
solute concentration outside the cell is less than
inside the cell water moves into cell and cell swells cell wall protects from lysis

hypertonic environments
inside water leaves the cell plasmolysis occurs

solute concentration outside the cell is greater than

Penicillin and lysozyme studies provide

evidence of role of cell wall, i.e. cells treated with both lyse in hypotonic solution
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Protoplast formation and lysis

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Components Outside of the Cell Wall

outermost layer in the cell envelope glycocalyx Capsules mainly polysaccharides,

provide protection against predators, chemicals and dessication Slime layers more diffuse than capsules, may aid mobility S layers
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aid in attachment to solid surfaces e.g., biofilms in plants and animals

Bacterial capsules

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Bacterial glycocalyx

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e.g. Glycocalyx Aerobic granular sludge in biofilms

Romain Lemaire

Glycocalyx in biofilms

S Layers
regularly structured layers of protein or
glycoprotein that self-assemble

Outside cell wall in G+ and membrane in G protect from ion and pH fluctuations, osmotic
stress, enzymes, and predation

maintains shape and rigidity promotes adhesion to surfaces protects from host defenses potential use in nanotechnology
S layer spontaneously associates
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S layer with distinct floortile pattern

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Archaeal Cell Envelopes

differ from bacterial envelopes in the molecular
makeup and organization S layer may be only component outside plasma
membrane some lack cell wall capsules and slime layers are rare

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Archaeal Membranes
composed of unique
lipids isoprene units (five
carbon, branched) ether linkages rather than ester linkages to glycerol

some have a

monolayer structure instead of a bilayer structure

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Archaeal membranes

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Archaeal Cell Walls Differ from Bacterial Cell Walls

lack peptidoglycan most common cell wall is S layer may have protein sheath external to
S layer

S layer may be outside membrane and

separated by pseudomurein

pseudomurein may be outermost layer

similar to gram-positive microorganisms

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Cell envelopes of Archaea

Methanospirillum Methanococcus, Halobacterium, Pyrodictium, Sulfolobus and Thermoproteus

Methanosarcina

Methanothermus and methanopyrus

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Metahnobacterium, Methanospaera, Methanobrevibacter, Halococcus and Natronococcus

Pseudomurein
NOTE: L-amino acids in linkers instead of Damino acids

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Bacterial and Archaeal Cytoplasmic Structures

Cytoskeleton Intracytoplasmic membranes Inclusions Ribosomes Nucleoid and plasmids

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Protoplast and Cytoplasm

protoplast is plasma membrane and
everything within

cytoplasm - material bounded by the

plasma membrane

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The Cytoskeleton
Internal architecture or scaffold of cell Not previously considered to be a part of
prokaryotes, but

homologs of all 3 eukaryotic cytoskeletal

elements have been identified in bacteria and 2 in archaea
Role in cell division, protein localization, and
determination of cell shape
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functions are similar as in eukaryotes

Bacterial cytoskeleton

e.g. FtsZ role in cell division e.g. Mbl maintains cell shape in rods, segregates chromosomes e.g. Crescentin induces curvature

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Organic inclusion bodies

glycogen
polymer of glucose units

poly--hydroxybutyrate (PHB)
polymers of -hydroxybutyrate Only found in prokaryotes Osmotically inert

Inorganic inclusion bodies

polyphosphate granules
also called volutin granules and metachromatic
granules linear polymers of phosphates

sulfur granules produced by sulphur bacteria using

H2S as energy source

magnetosomes
contain iron in the form of magnetite used to orient cells in magnetic fields Present in aquatic magnetotactic bacteria that want
to find nutrient rich waters!

Magnetosomes

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Microcompartments
not bound by membranes but
compartmentalized for a specific function
contain the enzyme ribulose-1,5,-

carboxysomes - CO2 fixing bacteria

bisphosphate carboxylase (Rubisco), enzyme used for CO2 fixation

Carboxysomes consist of polyhedral

shell

Shell prevents CO2 from escaping,

thus concentrating CO2
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Gas vacuoles
found in cyanobacteria and some other aquatic
procaryotes provide buoyancy aggregates of hollow cylindrical structures called gas vesicles Floating allows efficient capture of light for ATP production Vacuoles are aggregates of vesicles Vesicle walls are formed from proteins, which form a rigid cylinder impermeable to water but not gases Proteins can be collapsed to sink, assembled to float

Gas vacuoles and gas vesicles

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Ribosomes
complex structures entire ribosome
consisting of protein and RNA sites of protein synthesis bacterial and archaea ribosome = 70S eukaryotic (80S) S = Svedburg unit 16S molecule in small(i.e. 30S) subunit 23S and 5S in large subunit archaea has additional 5.8S in large one (also seen
in eukaryotic large subunit)

bacterial and archaeal ribosomal RNA

proteins vary

archaea (i.e. 68), more similar to eukarya (i.e. 78)

than to bacteria (i.e. 55)
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Bacterial ribosomes

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The Nucleoid
irregularly shaped region in bacteria and archaea usually not membrane bound (few exceptions) location of chromosome and associated proteins usually
a closed circular, double-stranded DNA molecule One copy of chromosome per cell

supercoiling and nucleoid proteins (HU) probably

aid in folding
nucleoid proteins differ from histones
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E.Coli nucleoids and chromosomes

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Plasmids
extrachromosomal DNA
molecules

found in bacteria, archaea, some fungi usually small, closed circular DNA

exist and replicate independently of

chromosome episomes may integrate into chromosome contain few genes that are nonessential confer selective advantage to host (e.g.,
drug resistance)
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Motility
Bacteria and Archaea have directed movement chemotaxis
move toward chemical attractants such as
nutrients, away from harmful substances

move in response to temperature, light, oxygen,

osmotic pressure, and gravity propeller

Flagellar movement - flagellum rotates like a Spirochete motility axial filaments flex and spin Twitching motility Gliding motility cells coast along solid surface
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Chemotaxis
movement towards a chemical attractant or away
from a chemical repellant

concentrations of chemoattractants and

chemorepellants detected by chemoreceptors on surfaces of cells

External structures: Pili and Fimbriae

fimbriae (s., fimbria); pili (s., pillus)
short, thin, hairlike, proteinaceous
appendages (up to 1,000/cell) mediate attachment to surfaces some (type IV fimbriae) required for motility or DNA uptake

sex pili (s., pilus)

similar to fimbriae except longer, thicker,

and less numerous (1-10/cell) genes for formation found on plasmids required for conjugation

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External structures: Flagella

threadlike, locomotor appendages extending functions
motility and swarming behavior attachment to surfaces may be virulence factors
outward from plasma membrane and cell wall

thin, rigid protein structures ultrastructure composed of three parts

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Patterns of Flagella distribution

Also, amphitrichous (one flagellum at each end of cell) and lophotrichous (cluster of flagella at one or both ends)

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Three Parts of Flagella

filament
extends from cell surface to the tip hollow, rigid cylinder composed of the protein flagellin some bacteria have a sheath around
filament

hook

links filament to basal body series of rings that drive flagellar motor
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basal body

Ultrastructure of Bacterial flagella

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Flagellar motor

Rotor

Stator

Archaeal Flagella
thinner more than one type of flagellin protein flagellum are not hollow hook and basal body difficult to
distinguish systems

more related to Type IV secretions growth occurs at the base, not the end
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The Bacterial Endospore

complex, dormant structure formed by some
bacteria

resistant to numerous environmental conditions

heat radiation chemicals Desiccation

Endospore resistance due to

Calcium complexed with dipicolinic acid Small acid-soluble DNA binding proteins Dydrated core Spore coat and exosporium protect
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Mature endospore

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Life cycle of an endospore

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Formation of Vegetative Cell

activation
prepares spores for germination often results from treatments like heating environmental nutrients are detected spore swelling and rupture of absorption of
spore coat loss of resistance increased metabolic activity

germination

outgrowth - emergence of vegetative

cell
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Germination
transformation of
endospore into vegetative cell complex, multistage process

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Comparison of Prokaryotic and Eukaryotic Cells

Take home messages:

Structural-conformation relationships key to
elucidating roles of intracellular molecules, and thus intracellular processes processes (e.g. in biofilm systems, current area of research at SCELSE here at NTU!)

Similar approach to elucidate extracellular