Kachur - Topic 2 - Bacterial Structure Flashcards
What is peptidoglycan?
- repeating disaccharide of NAG and NAM
What is NAG
- N-acetyl glucosamine
- a monosaccharide derivative of glucose
- amide between glucosamine and acetic acid
What is NAM
- N-acetyl muramic acid
- monosaccharide derivative of NAM
How are NAG and NAM cross-linked
with oligopeptides at the lactic acid residue of NAM
How do crosslinking peptides between gram positive and gram negative differ
Gram pos
- 9-15 aa
- often pentaglycine cross link
Gram neg
- 2 aa crosslink
- tetrapeptide links directly to another tetrapeptide
What is special about the D isomer of amino acids
- uncommon in nature
- used in bacteria to avoid protease digestion
- key to regulating disease
- connects through lysine component
What is N. gonorrhea?
- gram negative
- highly cross-linked -> harder to break down
- resistant to some antibiotics
What is the lipopolysaccharide layer?
- major component of the outer membrane of Gram negative bacteria
- elicit strong immune responses (O antigen can induce strong immune responses)
- LPS contribute greatly to the structural integrity of the bacteria and protecting the membrane from chemical attacks
How does LPS affect the charge of the cell membrane
- LPS also increases the negative charge of the cell membrane and helps stabilize the overall membrane structure
- LPS requires association with divalent cations (Mg++) to neutralize charge repulsion
What are the 3 molecular domains of LPS?
- lipid A
- the core
- the O-antigen
What are the non-essential components of the LPS
- the core and the O antigen
What is Lipid A?
- phosphorylated glucosamine disaccharide with associated fatty acids
- 4-6 fatty acids tails
- allows bilayer formation
- antigenic
- bacteria can function with only Lipid A
What is the core?
- attached directly to Lipid A
- mostly sugar but can contain amino acids
- diverse
What is Kdo?
the most common sugar in LPS
What is the O-antigen
attached to core olysaccharide
- highly diverse
- often determines strain type
What is the difference between rough and smooth LPS?
- rough O antigen generally associate with non-pathogens (less variable, usually less resistant to antibiotics)
- smooth can generally be more virulent than rough
- long O antigens are associated with virulence
What is the lipid A domain responsible for
toxicity of the gram negative bacteria
- upon bacterial lysis by the immune system, fragments containing lipid A are released into circulation causing:
- fever, diarrhea and endotoxic shock
What are the different types of PAMPS
- peptidoglycan
- lipid A
- teichoic acid
- capsules
- these are all unique to bacteria -> allow host detection
What are toll-like receptors (TLRs)?
- recognize PAMPs
- expressed in all cells of innate immune system (either on cell surface or endosomes)
- when a TLR is bound by a ligand, it activates a signalling cascade that initiates the release of pro-inflammatory cytokines
What is TLR6?
- recognizes peptidoglycan in gram +
- recognizes lipoteichoic acid in gram +
- recognize lipoproteins of gram -
What is TLR4?
- recognizes LPS in gram -
- found on cell surface
What is TLR3?
- found in endosomes
- detects intracellular + phagocytosed pathogens
What are NODs
- nucelotide-binding domain, leucine rich-repeat - containing receptors)
- sensors of intracellular PAMPS
- overlap between TLR cascades and NOD cascades
What is the pathogen profile of helicobacter pylori?
- gram negative
- acid tolerant
- infects lining of gut (asymptomatic)
- cause acute gastritis + chronic inflammation (ulcers)
- survives in the gut by secreting a base to neutralize acid + liquify mucus
- flagellar bundle: spirochaete shape allows it to penetrate mucus layer
What does NOD 1 recognize?
a modified dipeptide commonly found in gram negative
What does NOD 2 recognize?
peptidoglycan in both gram + and gram -
What is the pathogen profile of Listeria monocytogenes?
- gram negative
- causes listeriosis: serious infection usually caused by eating contaminated food
- likely to sicken pregnant women and their newborns, adults aged 65 or older and people with weakened immune systems
- can grow 0-37 degrees Celcius
- present in soft cheeses + raw milk
- attached to host via D-galactose (sugar on surface)
- can survive in phagocytes + can manipulate host cytoskeleton
How does Lm evade NLR recognition
50% of NAG replaced by glucosamine through deacetylation by the PgdA enzyme
What are some modifications in the cell envelope that help Lm evade NLR recognition
- Deacetylation of NAG by PgdA
- Glycosylation of teichoic acid = mechanism that enhances virulence is unknown
- Lysine is added to cell walls to increase + charge
- Adaptation for virulence => enhancing + charges which allows avoidance of cationic AMPs
Why are lipid A modifications important
essential to evade AMPs and TLR4
What are evasion strategies of Helicobacter pylori that promote survival in the host
- Dephosphorylation = removes (-) charge, resistant to CAMPs
- Removal of (-) charge by adding amine
- 2 fewer HC tails: hexacylated lipid A is highly antigenic
What are evasion strategies of Yersinia pestis that promote survival in the host
- when residing in the flea vector, Y. pestis produces an endotoxic hexa-acylated lipid A that is a strong immunostimulant in humans
- temperature change from fleas in humans causes changes in gene expression => turns on virulence genes
- in fleas = hexacylated lipid A is antigenic but not to flea
- Yersinia is virulent at 37 degrees
- enzyme is activated with temp shift that cleaves 2 acyl chains = TLR4 evasive
What are evasion strategies of Vibrio cholera
water borne illness that exists in copepod reservoirs
- addition of a diglycine to Lipid A decreases AMP sensitivity
What are antimicrobial peptides?
- potent members of the innate immune response
- have been shown to kill gram-negative and gram-positive bacteria
What are different subgroups of antimicrobial peptides?
- alpha helices
- beta sheets
- mixture of alpha and beta
- cyclic
- extended
What is Kalata B2?
- found in plants
- shown to have insecticidal, antitumor, antimicrobial
What are the three models of action mechanisms by AMPs
A) Barrel-Stave model: AMP inserts perpendicularly
B) Carpet model: small areas of membrane are coated in AMPs
C) Torodial pore model: represents Barrel Stave except AMP associates with phospholipid head