Lecture #2 - Cell Structure & Function PART 2 Flashcards
Peptidoglycan make up…
bulk but not all cell wall structure
Species of _____ separated into two groups based on Gram stain
Bacteria
Gram +
- still has a (-) cell surface
- THICK peptidoglycan
Gram -
- has net (-) charge
- THIN peptidoglycan
Some bacteria are exceptions being neither Gram + or Gram -
Mycoplasma
Gram-positives and gram-negatives have different cell wall structure
• Gram-negative cell wall
- Two layers: LPS (lipopolysaccharide) and peptidoglycan (THIN)
• Gram-positive cell wall
- One layer: peptidoglycan (THICK)
Cytoplasmic membrane
NOT a component of the cell wall
- just the boundary that forms around cytoplasmic content
- for all living cells
Peptidoglycan
RIGID layer that provides strength to cell wall
- organism can handle itself under extreme conditions since it’s just 1 cell
Describe difference b/t human cells and bacteria cells (in terms of cell wall)
- humans have NO cell wall, just have a plasma membrane (have lots of cells)
- BUT bacteria cells are unicellular, therefore anything that happens to that 1 cell will be the end of the cell structure
- so using peptidoglycan it has this peptidoglycan so it can better handle these situations/withstand so it’s not dying easily
Polysaccharide
peptidoglycan is formed by polysaccharide which is a complex sugar structure
Polysaccharide composed of:
- N-acetyl glucosamine (NAG) and N-acetylmuramic (NAM) acid
- Amino acids
- Lysine or diaminopimelic acid (DAP)
- Cross-linked differently in gram-negative bacteria and gram-positive bacteria
- Form glycan tetrapeptide
The Polysaccharide is…
crosslinks made out of amino acids
- link top to bottom, in order to make the structure of the peptidoglycan chain nice & strong
The L & D on the polysaccharide is…
indicative of the stoichiometry
- uses a D form of an AA that we don’t see in our cell
NAG-NAM-NAG-NAM-NAG-NAM-etc.
disaccharide is the first 2 pieces
- polymerizes again & again to create a chain around the cell
- comprises peptidoglycan layer
The AA chain only comes out of the…
NAM
Crosslinks in the peptidoglycan…
makes lattice structure strong, so if you apply a horizontal force of any kind, it will to an extent be able to withstand that
What would you prefer if given the option on the arrangement of the polysaccharide?
LEFT one –> requires less GTP
- b/c attaching AA’s by peptide bonds is a massive GTP requiring process (v. expensive) & you also need all the Gly residues
- *total for right one will need 5 additional gly residues & have to form additional 5 peptide bonds (so more energy)
What does penicillin target?
the enzyme used to form the polysaccharide
If you’re taking penicillin to target an infection, what part of the cell structure will NOT be okay? Chains or crosslinks that reinforce structure?
CROSSLINKS
If you look at a micrograph of bacterium that’s trying to grow in the presence of penicillin, you literally see holes blown on the side of the bacterium. Why would this organism have holes blown on the side as a result of having this cross link inhibition that penicillin provides?
NO STRENGTH
- & water wants to move in
- when that happens, the organism will have no protection b/c the crosslinks are being inhibited
Penicillin is useful against gram + organisms NOT against gram -? Why?
- more peptidoglycan –> more crosslink
- more repercussion for that type of organism
- b/c gram - some only have 1 PD layer so penicillin is essentially useless
Describe the structure of peptidoglycan
- NAG & NAM
- N-Acetyl group - acetyl group linked with a N to C2 of the sugar (true for NAM & NAG)
- Peptide cross-links - perpendicular
- B(1,4)
- Lysozyme-sensitive bond
- Glycan tetrapeptide - 4 AA’s attached to a sugar
Lysozyme-sensitive bond
incorporate it in sweat, saliva, & tears
- destroys NAM/NAG linkages specifically b/t NAM & NAG
- when it breaks that bond, the linear chains are fragmented (disassembling the cell wall of the organism)
- in doing so it protects our eyes, skin, mouth against bacterial infection
- but has to have peptidoglycan & will have more of an impact on organisms that are gram + b/c they have thick layers of PD
More than 100 different PG/PD structures identified…
- have same core PD/PG arrangement & crosslinks provided that they have more than 1 layer but way they form this will be diff. depending on the species
Vary in peptide cross-links and/or interbridge
No ______ is present in gram-negative Bacteria (e.g. E. coli). Why?
INTERBRIDGE
- b/c can have 1 single layer of PD (therefore, nothing to crosslink to)
The interbridge in Staphylococcus aureus (gram positive) is made up of…
5 glycine residues
Gram - have…
- NO interbridge
- can have crosslinks - but if it’s just 1 layer than not
- THIN PD
- if more than 1 layer of PD it wouldn’t have interbridge but still crosslinks
Glycosidic bonds b/t…
NAM & NAG
Peptide bonds…
AA’s used to form chains
Gram-positive cell walls
• Contain up to 90% peptidoglycan –> very THICK
- forms a helic (3D)
• Common to have TEICHOIC acids
- LIPOTEICHOIC ACIDS
Teichoic acids
(acidic substances) embedded in their cell wall
- common in gram + cell walls
- like toothpick that puncture bread (PM) underneath
Lipoteichoic acids
teichoic acids covalently bound to membrane lipids
- toothpick penetrates through bread too (PM) underneath
Teichoic acids & Lipoteichoic acids…
DO NOT exist in gram - organisms which have LESS PD
Gram + only (i.e. may not be present, but if is will be in gram +)
1) Lipoteicholic acids
2) Endospores
3) Interbridges
Model of Peptidoglycan Surrounding the Cell
- Backbone formed of NAM and NAG connected by GLYCOSIDIC BONDS
- Crosslinks formed by PEPTIDES
• Peptidoglycan strand is HELICAL
- Allows 3-DIMENSIONAL CROSSLINKING
- E. coli has 1 layer
- Some cell walls can be 50 - 100 layers thick, e.g. BACILLUS SPECIES
Peptidoglycan strand is helical, allows…
Allows 3-dimensional crosslinking
E. coli has ___ layer
1
Some cell walls can be ____ layers thick, e.g. _______
50-100
Bacillus species
Bacillus species
- cell walls is 50 - 100 layers thick
- gram +
- purple
- they form endospore structures (indicator of gram +)
Prokaryotes that lack cell walls
Mycoplasmas
Mycoplasmas
• Prokaryotes that LACK cell walls (comparable to animal cell)
- v. small
- Group of pathogenic bacteria
- Have sterols in cytoplasmic membrane–adds strength and rigidity to membrane
Mycoplasmas have sterols in cytoplasmic membrane…
– adds strength and rigidity to membrane
sterols - cholesterol/other sterols which adds hot/cold stability (we also use cholesterol)
- better able to withstand temp flex’s
- usually a eukaryotic thing
Thermoplasma
- Prokaryotes that lack cell walls
- Species of Archaea (would never have peptidoglycan anyway - no cell wall here!)
- Contain LIPOGLYCANS in membrane that have strengthening effect
Gram negatives cell wall…
- total cell wall contains ~10% peptidoglycan (THIN)
* Most of cell wall composed of outer membrane, aka lipopolysaccharide (LPS) layer (still a bilayer)
Most of GRAM - cell wall composed of outer membrane, aka lipopolysaccharide (LPS) layer
- LPS consists of CORE POLYSACCHARIDE and O-POLYSACCHARIDE
- LPS replaces most of phospholipids in outer half of outer membrane
- ENDOTOXIN (lipidA): the toxic component of LPS
Endotoxin
toxic when RELEASED from dead cell (not toxic when attached)
Endotoxin (lipid A) is only in…
gram -
What does the outer leaflet of the outer membrane consist of?
- o-specific polysaccharide
- core polysaccharide
- Endotoxin (lipid A)
(LPS & phospholipids basically)
Gram (-) sepsis
- organism that’s everywhere growing & multiplying
- endotoxin dramatically activates the immune system b/c it’s gram -
- immune system does freak out & causes blood vessels to dilate (inflammation - okay in foot or throat b/c increases flow when you dilate a blood vessel & allows delivery of O2 & nutrients & immune cells for healing, immune responses to clear things out)
- BUT when its sestemic (sepsis), that inflammatory response causes all of arterial to vasodilate
Why might this be a problem (gram - sepsis) if all BV’s are dilating not just ones located in an infected tissue?
- BP drops dramatically LOW –> DON’T have enough pressure to force fluid to your tissues to deliver O2 & nutrients
- tissues start to die
- amputation b/c wasn’t enough pressure to feed the tissues
(like frost bite - vasoconstriction)
Why should you NEVER give an antibiotic that aggressively kills bacteria if you have sepsis?
b/c ENDOTOXIN gets RELEASED when you aggressively kill bacteria in really high [ ] in the cell structure
- can kill them
What should you use instead for gram - sepsis?
would want to use a BACTERIOSTATIC drug - keeps everything constant
- should be used to treat gram - sepsis b/c it interfers with growth
- prevent them from increasing # & immune system goes & cleans up the organisms that we’re there
- drug doesn’t do killing & need to have a functional immune system to do the destruction
Porins in the LPS/the outer membrane are…
NON-SPECIFC
- just providing a channel for polar material that’s size appro.
Things like ABC transport, group translocation etc. in the LPS/the outer membrane are…
HIGHLY specific
- allow organisms to enter through
Bacteriostatic drug should be used to treat…
gram - sepsis
Cytoplasmic membrane
highly specific with transmem. proteins that select for CERTAIN nutrients or ions to be able to enter the cell
LPS: The Outer Membrane
Periplasm:
space located between cytoplasmic and outer membranes
• ~15 nm wide
• Contents have gel-like consistency –> VISCOUS (lots of nutrients)
• Houses many proteins
ex: periplasmic binding proteins
- bring nutrients to the ABC transporters
think: yard
- just came through fence & is able to wonder around & if you meet specificity of the periplasmic proteins you can enter house (cytoplasmic envir. of cell)
Ex of proteins in the periplasm
ex) periplasmic binding proteins
- bring nutrients to the ABC transporters
LPS: The Outer Membrane
Porins:
channels for movement of hydrophilic low-molecular-weight
substances
- have Beta pleated sheets (unique secondary structure)
- non-specific entry
- water-soluble & hydrophilic, but small enough to meet size of the pore
- therefore, doesn’t serve as permeability barrier
LPS: The Outer Membrane
Porins:
channels for movement of hydrophilic low-molecular-weight
substances
- non-specific entry
- water-soluble & hydrophilic, but small enough to meet size of the pore
- therefore, doesn’t serve as permeability barrier
Porins have…
Beta pleated sheet (unique secondary structure)
Relationship of Cell Wall Structure to Gram stain
• In Gram stain reaction, insoluble crystal violet-iodine (CV-I) complex forms inside cell
- crystal violet sticks to (-) charge of cell
- iodine bulkens/increases structural size of the dye
• Complex is extracted by alcohol (decolourizing step) from gram-negative, not gram- positive bacteria
Reason:
• Gram-positive bacteria have thick cell walls consisting mainly of peptidoglycan
- Becomes dehydrated during alcohol step so pores in wall close
- Prevents CV-I complex from escaping –> cell remains purple
• Gram-negative bacteria – alcohol penetrates OM
- CV-I extracted from cell
- Cells appear nearly invisible (colourless) until counterstained with second dye (safarin - basic pink dye, stick to everything (-) charged (including gram +’s) but purple overpowers them)
- but now includes gram -‘s that were colourless, turning them pink
Archaeal Cell Walls
- NO peptidoglycan (prokaryotes)
- Typically NO outer membrane
- Pseudomurein
- Cell walls of some Archaea LACK pseudomurein
- S-Layers
Prokaryotes NEVER have…
peptidoglycan
Pseudomurein
Archeal don’t have PD but still have a cell wall:
• Polysaccharide similar to peptidoglycan
- not common among all archaea but some have it
• Composed of N-acetylglucosamine (NAG) and N-acetyltalosaminuronic acid (NAT) (REPLACES NAM of PD)
• Found in cell walls of certain methanogenic Archaea
- makes CH4 (methane)
Pseudomurein structure
- NAG & N-Acetyl-talosaminuronic Acid
- Lysozyme-INsensitive B (1,3)
- Peptide cross-links (b/t NAG & T for these - PD was b/t NAM residues M—M)
Describe B (1,3) lysozyme-INsensitive in Pseudomurein
NOT B (1,4), therefore WON’T work as a lysozyme substrate
- therefore, a lysozyme that we have in our sweat, saliva & tears WOULDN’T be effective agains the pseudomurein cell wall b/c it doesn’t have the B (1,4) linkage present b/t the NAM & NAG that was discussed for PD
- a lysozyme b/c it’s an enzyme & highly specific, WOULDN’T be able to cleave this & wouldn’t have effectiveness against the cell wall structure
B/c of the B (1,4) linkage, if archaea ever became pathogenic (b/c it can happen in future)…
our own defences in our sweat, saliva & tears WOULDN’T be effective in order to disturb the cell wall so we would have to rely on other means
Archaeal Cell Walls
S-Layers
• Most COMMON cell wall type among Archaea
- numerically dominants the pseudomurein
• Consist of protein or glycoprotein (protein that has sugar groups attached to the structure in order to create an alternative function)
• Paracrystalline structure
- crystallized structure –> ordered, firm
- SOME Archaea have ONLY S-layer (no other cell wall components)
- MOST have ADDITIONAL cell wall elements
Summary of Archaeal Cell Walls
- Variety of structures possible (diversity)
- SOME (pseudomurein) closely resemble peptidoglycan
• Others LACK polysaccharide (long chain of sugar - NOT just sugar units that are strategically placed onto a protein) completely
ex) S layer (protein or glycoprotein)
• Most Archaea contain some type of cell wall structure –
functions to prevent osmotic lysis and give shape (coccus, bacillus, etc.)
- EXCEPTION: thermoplasma spp. - archaea that has no cell structure
• Because they lack peptidoglycan, Archaea are resistant to lysozyme and penicillin
i.e. can’t use penicllin to target
How do Archaeal cell wall vary?
- pseudomurein, S-layer, add. components that could be added etc.
- compared to PD which is consist for almost all bacteria
Which Archaeal cell walls lack polysaccharide completely?
ex) S layer (protein or glycoprotein)
What is an exception to, “Most Archaea contain some type of cell wall structure?”
exception: thermoplasma spp. - Archaea that has NO cell structure
Because they lack peptidoglycan, Archaea are resistant to lysozyme and penicillin (explain)
i.e. can’t use penicillin to target
penicillin is not an effective chose B/C it TARGETS PD in partic., if you did gain an archeal infection when that would become pathogenic 1 day
- b/c it doesn’t have PD & PD crosslinks are specifically gonna be targeted by the penicillin drug
- therefore, can’t use penicillin to treat an archeal infection ever, b/c the target for that drug isn’t present in the archeal cell wall structure or in the archeal cell wall at all
- so you would need to use a protein syn. inhibitor for ex.
Cytoplasm
material bounded by plasma membrane (PM)
=CYTOSOL (FLUID) & ALL COMPONENTS WITHIN
EX) RIBOSOMES
Protoplast
PM + CYTOPLASM and everything within
• Macromolecules – amino acids, nucleotides, etc
• Soluble proteins ex) enzymes for glycolysis - allow metabolic pathways to happen with ease & simply b/c the enzymes can shift & find substrate & move things around
• DNA and RNA (nucleoid)
- RNA: transcripts produced for protein synthesis i.e. make a protein from a transcript thats come out
Proteins
• Serve many functions:
• Enzymes – Catalyze chemical reactions (rapid & efficient)
• Transport proteins – Move other molecules across membranes (that’s polar & of a partic. characteristic)
ex) porins, ABC transporters etc.
- Structural proteins – Help determine shape of the cell (morphological det)
- Involved in cell division
• Proteins are made of polypeptides
- think: hemoglobin - come together properly for function
• Polypeptide – a long polymer of amino acids joined by peptide bonds
Proks can have a cytoskeleton…
made out of protein (proteination - made out of protein, plays role in determining shape or morphology of cell), the same way euk’s have a cytoskeleton
Describe the peptide bonds of proteins
- partial double bond characteristics
- restricts rotation (shorter)
- plays role in determining shape that proteins can assume
The Nucleoid
• Region that contains the genome
- NOT a fenced off area but good chance you’ll find a chromosome there
- DNA
- Carries genetic info of all living cells
- Polymer of deoxyribonucleotides
The Nucleoid
The typical bacterial genome:
Single circular double stranded (ds) DNA chromosome (ALWAYS carries the genetic info in DNA b/c it’s a LIVING cell (prok, animal or plant cell)
MAY have one or more plasmids
• Smaller circular dsDNA
The typical bacterial genome, MAY have one or more plasmids:
- Smaller circular dsDNA
- Self-replicating –> autonomously!
- can photocopy themselves when rest of cell & chromosome are not duplicating & div.
- these bad bacteria hand out these copies v. generously to other bacteria & pass it along to another (to make someone sick) –> v. generous & like to share
- how antibiotics resis. is able to spread throughout community
• Carry non-essential genes
- BUT if an organism finds itself in a partic. envir. or under a partic. set of conditions, it’s def gonna BENEFIT from having them
- Selective advantage –> allow cell to produce a toxin or enzyme or a gene for penicillin resistance to allow a cell to survive presence of penicillin b/c it gives it the phenotype of resistance to that partic. drug
- if there, it will benefit from it - like ripping recipes that we don’t use
- Ex) Genes for antibiotic resistance
Genome =
TOTAL content of DNA in the cell!
If the plasmid was in the cell, would it be included in the genome?
YES
Is it guaranteed that the plasmid would be included in the cell tomorrow? Or can cell kick it out if don’t need?
NO it’s not guaranteed - it can come & go
- cells are v. genetically plastic (pick up/get rid of/moving around cellular comp. constantly)
What does the Genome include?
ESSENTIAL GENES
including: enzymes for glycolysis, ETC protein, transport proteins in mem. for ex
cell CANNOT live without - foundational for life & survival
What does the Plasmid include?
NON-ESSENTIAL GENES
- don’t need them - not necessary for energy production transporting nutrients in cell, but if cell has these plasmids it provides some kind of a selective advantage
Ribosomes in prok
Site of protein synthesis
70S ribosome (SMALLER than 80S euk)
• 2 parts
- 30S subunit (Small subunit) SSU
- Protein
- 16S rRNA - 50S subunit (large subunit) LSU
- Protein
- 23S and 5S rRNA
Cytoplasmic ribosomes
• Cytoplasmic proteins
PM associated ribosomes
• Membrane proteins
• Proteins to be exported from the cell
70S prok ribosome is _______ than 80S euk
SMALLER
What does the 16S, 23S or 5S rRNA mean?
how many nucleotides will comprise the piece
What ribosomes can a prok. ribosome have?
70S (50S + 30S)
& also could have 70S bound to PM - NOT all prok’s have but if there, they make protein that get exported (like rough ER) to outside of cell - could be toxin, signalling molecule that communicate with buddies nearby etc.
What are 2 differences b/t 23 rRNA & 16S rRNA?
1) 23 rRNA is LARGER - longer length
2) Nucleoid sequence is DIFF.
Ribosomes..
take a messenger rRNA piece & convert it to protein like a factory
- assembles AA’s forming peptide bonds by looking at what template is asking for in messenger rRNA, forming a polypeptide & then repeat
What ribosomes can a Euk cell have?
1) 80S ribosomes (40S + 60S)
2) Euks have 70S ribosomes too in mitochondria!
Do Euk’s have 70S CYTOPLASMIC ribos?
NO - b/c cytoplasmic ribo’s are 80S!
they do have 70S ribosomes but they are in the mitochondria!
Are Cell Surface Structures present on all bacteria or just some? Where are they?
Present on SOME bacteria
OUTSIDE of CELL WALL (1st thing you’ll see)
List the possible Cell Surface Structures that can be present on SOME bacteria:
- Capsules and slime layers
- Fimbriae
- Pili
List the Cell Surface Structures present on SOME bacteria:
- Capsules and slime layers
- Fimbriae
- Pili