Chapter 4 Part 3 The Cell Wall of Prokaryotes: Peptidoglycan and Related Molecules Things to look up • General structure of sugars – How do sugars bind together? – What is the difference between a and bglycosidic linkages? • General structure of amino acids • Peptide bonds – What 2 groups link together 2 amino acids? Cell Walls • Eukaryotes – plants; differ chemically from prokaryotes; simplier in structure and less rigid • Bacteria – Peptidoglycan – Destroyed by lysozyme – cell lysis • Archae – lack peptidoglycan but contain walls made of other polysaccharides or protein Functions of the Cell Wall • Support/cell shape • Surrounds the plasma membrane and protects it and the interior of the cell from adverse changes in the outside environment • Prevents cells from rupturing • Point of anchorage for flagella Functions of the Cell Wall • Contributes to the ability of bacteria to cause disease – Important in the attachment to host cells – Barrier to some molecules • Site of action of some antibiotics • Chemical composition of the cell wall differentiates gram + from gram - bacteria Site of action of some antibiotics • Why is it important that antibiotics work on the cell wall? – Eukaroytes (besides plants) do not have cell walls – If kill bacteria, human cells will still live Peptidoglycan • Found in both gram + and gram – bacteria • Consists of a sugar backbone • Different chains of these sugars are linked together by peptide bonds between amino acids Peptidoglycan • Consists of a sugar backbone of alternating repeats of Nacetylglucosamine (NAG, G) and N-acetylmuramic acid (NAM, M) • NAM and NAG in b1-4 glycosidic linkage • NAM is cross-linked between strands by short peptides attached to NAM Peptidoglycan • Consists of four amino acids (peptides) • L-alanine, D-glutamic acid, either lysine or diaminopimelic acid (DAP) and D-alanine • Alternating pattern of L and D amino acids – Unique because it is always L-amino acids found in other proteins 4 amino acids in peptidoglycan linked to NAM different for Gram + and Gram - How do the different chains of peptidoglycan link together? • Different for gram + and gram – bacteria • Gram – Linkage of amino group of DAP and carboxyl group of terminal D-alanine group How do the different chains of peptidoglycan link together? • Gram + – Peptide interbridge – Kinds and number of amino acids vary Peptidoglycan • Gram + - thick layer of peptidoglycan • Gram - - thin layer of peptidoglycan • Basis for the gram stain Bacterial Cell Wall Types • Gram type describes the structure of cell wall which influences the way it stains • Thicker peptidoglycan holds crystal violet – Gram + • Counterstain is pink – Gram -; thinner peptidoglycan Gram + bacteria very sensitive to the action of penicillin and lysosome • Penicilin interferes with the final linking of the peptidoglycan rows by a peptide cross-bridge • Lysozyme is an enzyme found in tears and saliva that breaks the b-1,4-glycosidic bonds between NAM and NAG • Gram + cell wall is mostly peptidoglycan so it is more sensitive than gram – cell walls Gram - vs. Gram + • Gram-negative Bacteria have only a few layers of peptidoglycan • Thin peptidoglycan • Only about 10% of the cell wall is peptidoglycan • 2 layered membrane • Periplasm between the two layers Gram + • Thick peptidoglycan • (90%) • negatively charged teichoic acid • Cross-linking occurs with a peptide interbridge (amino acids involved differ) Gram positive –Teichoic acid • Polymer of glycerol or ribitol • Joined by phosphate groups • Amino acids are attached Gram positive –Teichoic acid • Lipoteichoic acid (lipid + teichoic acid) – Spans the peptidoglycan layer and is linked to the plasma membrane • Teichoic acid – Linked to the peptidoglycan layer • Purpose: stability, passage of ions, gives negative charge to the cell Teichoic acid is negatively charged • Bacteria are stained with + dyes Gram - bacteria • Contain an inner and outer membrane • Peptidoglycan in between – thin layer – bound to lipoproteins in the outer membrane • Outside the cytoplasmic membrane is the periplasmic space, a fluid filled space – Contains degradative enzymes and transport proteins – Proteins transported here by the SecYEG system Functions of the outer membrane of gram - bacteria • Gives a negative charge to the cell • Important in evading phagocytosis and host defense • Pathogenic properties • Selective barrier – Pore for entrance of hydrophilic molecules – Barrier to certain antibiotics and digestive enzymes Outer membrane of gram - bacteria • LPS • Lipoproteins – Anchor to the peptidoglycan • Porins – Proteins that form pores (channels) in the outer membrane – Wide enough to allow passage of small hydrophilic molecules – Large hydrophobic molecules cannot penetrate LPS = lipopolysacharide • lipid A – endotoxin properties, which may cause violent symptoms in humans – Anchor to the membrane • a core polysaccharide – 6 or 7 C- sugars (Gal, Glu, NAG, Ketodeoxyoctonate or KDO, etc.) • O-specific polysaccharide – 6 C- sugars (Gal, Glu, Man, Rhm, etc.), repeating units of 45 sugars, often branched – Reaches out into the environment – Function as antigens – differentiate different bacteria What does LPS do? • Activate Toll receptors Different Toll receptors for different pathogens • Toll receptors part of innate immunity • Some receptors are extracellular while some are intracellular • Some receptors dimerize Gram Stain • The structural differences between the cell walls of gram-positive and gram-negative Bacteria are thought to be responsible for differences in the Gram stain reaction • Alcohol can readily penetrate the lipid-rich outer membrane of gram-negative Bacteria and extract the insoluble crystal violet-iodine complex from the cell Gram stain • http://www.youtube.com/watch?v=sxa46xK fIOY&list=PLrAEgIY86I6wYIgx3iEKvyaRFzwuuixr&index=13 Gram Stain Some organisms have no cell walls • Mycoplasma – Intracellular parasite – Can only survive inside of their host – no need for cell wall but have tough membranes • More resistant to rupture than other bacteria – Another difference from other bacteria is that mycoplasma contain sterols that help protect from lysis Archaea have unusual cell walls • No peptidoglycan • Typically no outer membrane • Pseudomurein – Polysaccharide similar to peptidoglycan – Composed of Nacetylglucosamine and Nacetylalosaminuronic acid Archaea have unusual cell walls • Thermoplasma has no cell wall (extremely stable lipid membrane) • S-Layers – Most common cell wall type among Archaea – Consist of protein or glycoprotein