SBS-713

Archaea, the third domain of life

Differences Among Bacteria, Arcahea, and

Eukarya
How Archaea cope with extreme environments

Dr. Mehwish Aslam Lecture 2

Last lecture
• Start of life on the Earth
• Diversification of life
• Origin of Archaea
• Type of Archaea
• Timeline …
Genome of the Archaeon Methanococcus
jannaschii was sequenced in 1996.
Sequencing of M. jannashcii confirmed
Carl Woese’s long-standing hypothesis that
life traces back to three main lineages, one
of which (Archaea) includes prokaryotes
that share a more recent common
ancestry with eukaryotes than with the
prokaryotic “true bacteria”
What makes Archaea different from Bacteria and Eukarya
?
What makes Archaea survive in extreme environments
?
Archaeal Cell Wall
• There is no cell wall structure unique to all Archaea
•Different building blocks found in different archaeal clades
•Some Archaea do not contain cell walls, e.g.. Thermoplasma
Archaeal Cell Wall
Pseudomurein (Pseudopeptidoglycan)
• Shares structural similarities with bacterial murein yet presents significant
differences
• Pseudomurein usually consist of N-acetyl-L-talosaminuronic acid linked via a
β1-3 linkage to N-acetyl-D-glucosamine, unlike murein, which consists of
alternating N–acetylmuramic acids linked via β1-4 linkage to GlcNAc.
• The glycan strands of pseudomurein are cross-linked by peptides composed
of L-amino acids (glutamic acid, alanine, and lysine), (D-amino acids used in
murein).
• A novel biosynthesis pathway for pseudomurein has been proposed that is
not present in bacteria
Archaeal Cell Wall
Pseudomurein (Pseudopeptidoglycan)

Albers and Meyer, 2011 Essentials of Glycobiology [Internet]. 3rd edition.

Archaeal Cell Wall
S-Layer
• Archaea have proteinaceous cell wall, the S-layer, comprising a regularly
structured two-dimensional array based on a single protein species, the S-
layer glycoprotein, or a limited number of proteins.
• S-layer (surface layer) is a part of the cell envelope found in almost all archaea
• S-layers have a thickness between 5 and 25 nm.
• Ignicoccus spp. represent another interesting exception in the archaeal
domain in that they are the only species that possess two membranes and no
S-layer
Archaeal Cell Wall
S-Layer (a) Hydrophobic transmembrane domains
(b) Lipid-modified glycoprotein subunits
(c) Rigid wall layer (e.g. pseudomurein in
methanogenic organisms) as intermediate
layer between the plasma membrane and
the S-layer
(d) S-layer (glyco)proteins are bound to
the rigid peptidoglycan-containing layer
(e) S-layer is closely associated with the
lipopolysaccharide of the outer
membrane.

Sleytr et al. 2014

Archaeal Cell Wall
Proteinaceous Sheaths
• Rod-shaped cells of Methanospirillum hungatei and Methanosaeta
concilii form long filamentous chains in which each cell is surrounded by an S-
layer presenting hexagonal symmetry or by a rigid granular layer similar to an
S-layer
• Filamentous chains are further enclosed by a tubular proteinaceous sheath
• Based on several cross-links involving cysteines, these sheaths are highly
stable against proteases and detergents
• Proteinaceous sheaths can be Post-translationally modified by the attachment
of various sugars.
Proteinaceous Sheaths
 Archaeal Cell Wall
Halomucin
• Extremely halophilic Haloquadratum walsbyi has a unique square shape with
a length of 1.5–11 µm but a thickness of only 0.1–0.5 µm
• These cells are surrounded either by an S-layer or by two S-layer sheets
• These cells also secrete halomucin, an extremely large glycoprotein that is
highly similar to mammalian mucin, which stays loosely connected to the cell.
• Halomucin is heavily glycosylated, containing
more than 280 potential N-glycosylation sites
• This unique protective cell envelope allows H.
walsbyi to cope with salinities

Haloquadratum walsbyi
Archaeal Cell Wall
Halomucin
Archaeal Cell Wall
Glutaminylglycan
• The cell wall of the highly halophilic and alkaliphilic genus Natronococcus (3.5
M salt and pH 9.5–10) consists of a glutamine polymer.
• In contrast to poly-γ-D-glutamyl polymers present in
bacteria Bacillus, Sporosarcina, or Planococcus, the archaeal polymer is
formed from L-glutamines linked via the γ-carboxylic group, yielding a chain of
about 60 monomers.
• Archaeal poly-γ-L-glutamine chain is glycosylated, containing two types of
oligosaccharide in comparison to bacterial polymer.
• The first oligosaccharide consists of a GlcNAc pentasaccharide at the reducing
end and multiple GalA residues at the nonreducing end
• The second presents a GalNAc disaccharide at the reducing end and two Glc
units at the nonreducing end.
Archaeal Cell Wall poly-γ-L-glutamine chain
Glutaminylglycan

2nd Glycosylation 1st Glycosylation

Kandlera and Konig, 1998

Archaeal Cell Wall
Heteropolysaccharides
•Halococcus morrhuae
• An extreme halophile surrounded by an electron-dense 50–60-nm-thick
cell wall composed of a complex, highly sulfated heterosaccharide
consisting of glucosamine, galactosamine, gulosaminuronic acid, glucose,
galactose, mannose, glucuronic acid, galacturonic acid, N-acetylated amino
sugars, and sulfated subunits.
•Different heteropolysaccharides are thought to be connected via
glycine bridges between the amino groups of the glucosamines and
the carboxyl groups of the uronic residues.
Archaeal Cell Wall
Methanochondroitin
• Individual cells of Methanosarcina rely on an S-layer as their cell wall.
• A cubic aggregate of four cells (Sarcina) is covered by an additional rigid
fibrillar polymer called methanochondroitin.
• Degradation of methanochondroitin results in disaggregation of the cells,
underlining that the matrix is responsible for maintenance of the aggregate.
• Methanochondroitin, which is similar to eukaryotic connective tissue
chondroitin, is composed of a repeating trimer of uronic acid and two GalNAc
residues.
• unlike chondrotin, methanochondroitin is not sulphated, and the molar ratio of
GalNAc:GlcA is 2:1, not 1:1 as in chondroitin.
• Methanosarcina species can further modify the methanochondroitin
condition largely through the addition of glucose and galactose acids.
Archaeal Cell Wall
Methanochondroitin
Archaeal Cell Wall
Lipoglycan
• Members of the thermoacidophilic order Thermoplasmatales (pH 1–2 and
∼60°C), such as Ferroplasma acidophilum and Thermoplasma acidophilum,
lack a rigid cell envelope.
• Such organisms display a pleomorphic shape, similar to mycoplasma.
• Stabilization of the cell is most likely realized by the oligosaccharide portions
of lipoglycans and membrane-associated glycoproteins.
• The outwardly oriented glycan chains form a protective slime coat called the
glycocalyx.
 Archaeal Cell Wall
Glycocalyx
• A highly-hydrated fibrous meshwork of carbohydrates that projects out and
covers the membrane of endothelial cells, many bacteria and other cells.
• Proteoglycans and glycoproteins are generic structural components of
a glycocalyx
• Mediates cell attachment, retains humidity during exposure to dry
environments, protects against molecular and cellular antibacterial agents
• Can be present in form of
• Capsule – highly organized, tightly attached
• Slime layer – loosely organized, and attached
CM, cytoplasmic membrane; GC, glycocalyx; HP,
heteropolysaccharide; MC, methanochondroitin matrix;
OM, outermost membrane; PM, pseudomurein; PS,
proteinaceous sheath; SL, S-layer.
Cell membrane – Archaea and Bacteria

• Bacterial and eukaryotic membranes have

lipids with fatty acids connected to glycerol
by ester linkages.
• Archaeal membranes have lipids with fatty
acids linked to glycerol by ether linkages.
The Diversity of N-Linked Glycans in Archaea

• Archaea S-layer glycoproteins (and indeed, their other glycoproteins,

such as archaellins) bear N-linked glycans that present wider diversity in
terms of size, degree of branching, identity of the linking sugar,
modification of sugar components by amino acids, sulfate, and methyl
groups, and the presence of unique sugars than reported to date in
Bacteria or Eukarya.
Effect of antibiotics directed against cell wall synthesis

• Cell wall polymers of the various archaeal lineages are chemically

unrelated and differ considerably from the bacterial peptidoglycan
(murein), no common target for antibiotics directed against the
cytoplasmic steps of the cell wall synthesis of all prokaryotes is to be
expected.
• Classical antibiotics directed against murein biosynthesis, such as
fosfomycin, vancomycin and β-lactam antibiotics, have no growth
inhibiting effect against archaea
• The only known exception is Methanococcus vannielii, which is sensitive
to D-cycloserine, although this organism possesses no known D-alanine-
containing structure, the well-known target of D-cycloserine inhibition.
(mechanism not clear)
Effect of antibiotics directed against cell wall synthesis

• Antibiotics interfering with the lipid cycle, such as bacitracin and

gardimycin, are also inhibitory against different archaea, regardless of
whether they possess pseudomurein or other types of cell wall
polymers.
• These antibiotics may interfere with different lipid bound precursors of
various carbohydrate-containing polymers (pseudomurein,
heteropolysaccharides, glycoproteins) or with the biosynthesis of
isoprenoid diether lipids typical of archaea
• Tunicamycin, which inhibits the transfer of N-acetylglucosamine
residues in glycoprotein and murein biosynthesis, also prevents the
growth of Methanobacterium thermoautotrophicum.