Peptidoglycan

{{Short description|Polymer in bacterial cell walls}} {{Distinguish|glycopeptide|proteoglycan|glycoprotein}} '''Peptidoglycan''', '''murein''' or '''mucopeptide''' is a unique large macromolecule, a polysaccharide, consisting of sugars and amino acids that forms a mesh-like layer (sacculus) that surrounds the bacterial cytoplasmic membrane.<ref name="Madigan-2015">{{Cite book |last=Madigan |first=Michael T. |title=Brock Biology of Microorganisms |last2=Martinko |first2=John M. |last3=Bender |first3=Kelly S. |last4=Buckley |first4=Daniel H. |last5=Stahl |first5=David A. |publisher=Pearson Education Limited |year=2015 |isbn=978-1-292-01831-7 |edition=14 |location=Boston |pages=66–67}}</ref> The sugar component consists of alternating residues of β-(1,4) linked ''N''-acetylglucosamine (NAG) and ''N''-acetylmuramic acid (NAM). Attached to the ''N''-acetylmuramic acid is an oligopeptide chain made of three to five amino acids. The peptide chain can be cross-linked to the peptide chain of another strand forming the 3D mesh-like layer.<ref name="Madigan-2015" /><ref>{{Cite web |date=20 March 2011 |title=Animation of Synthesis of Peptidoglycan Layer |url=http://pharmaxchange.info/press/2011/03/animation-of-synthesis-of-peptidoglycan-layer/ |website=PharmaXChange.info |vauthors=Mehta A}}</ref> Peptidoglycan serves a structural role in the bacterial cell wall, giving structural strength, as well as counteracting the osmotic pressure of the cytoplasm. This repetitive linking results in a dense peptidoglycan layer which is critical for maintaining cell form and withstanding high osmotic pressures, and it is regularly replaced by peptidoglycan production. Peptidoglycan hydrolysis and synthesis are two processes that must occur in order for cells to grow and multiply, a technique carried out in three stages: clipping of current material, insertion of new material, and re-crosslinking of existing material to new material.<ref>{{Cite journal |vauthors=Belgrave AM, Wolgemuth CW |date=June 2013 |title=Elasticity and biochemistry of growth relate replication rate to cell length and cross-link density in rod-shaped bacteria |journal=Biophysical Journal |volume=104 |issue=12 |pages=2607–2611 |bibcode=2013BpJ...104.2607B |doi=10.1016/j.bpj.2013.04.028 |pmc=3686348 |pmid=23790368}}</ref>

The peptidoglycan layer is substantially thicker in gram-positive bacteria (20 to 80 nanometers) than in gram-negative bacteria (7 to 8 nanometers).<ref>{{Cite web |date=18 March 2016 |title=Bacteria |url=https://basicbiology.net/micro/microorganisms/bacteria |publisher=Basic Biology |vauthors=Purcell A}}</ref> Depending on pH growth conditions, the peptidoglycan forms around 40 to 90% of the cell wall's dry weight of gram-positive bacteria but only around 10% of gram-negative strains. Thus, presence of high levels of peptidoglycan is the primary determinant of the characterisation of bacteria as gram-positive.<ref>{{Cite encyclopedia |title=Bacteria |encyclopedia=Encyclopedia of Earth |publisher=National Council for Science and the Environment |location=Washington DC |url=https://editors.eol.org/eoearth/wiki/Bacteria |date=12 October 2014 |vauthors=Hogan CM |veditors=Draggan S, Cleveland CJ}}</ref> In gram-positive strains, it is important in attachment roles and serotyping purposes.<ref name="Salton-1996">{{Cite book |title=Structure. ''In:'' Baron's Medical Microbiology |vauthors=Salton MR, Kim KS |publisher=Univ of Texas Medical Branch |year=1996 |isbn=978-0-9631172-1-2 |editor-last=Baron S |edition=4th |chapter=Structure |pmid=21413343 |display-editors=etal |chapter-url=https://www.ncbi.nlm.nih.gov/books/NBK8477/}}</ref> For both gram-positive and gram-negative bacteria, particles of approximately 2 nm can pass through the peptidoglycan.<ref>{{Cite journal |vauthors=Demchick P, Koch AL |date=February 1996 |title=The permeability of the wall fabric of Escherichia coli and Bacillus subtilis |journal=Journal of Bacteriology |volume=178 |issue=3 |pages=768–773 |doi=10.1128/jb.178.3.768-773.1996 |pmc=177723 |pmid=8550511}}</ref>

It is difficult to tell whether an organism is gram-positive or gram-negative using a microscope; Gram staining, created by Hans Christian Gram in 1884, is required. The bacteria are stained with the dyes crystal violet and safranin. Gram positive cells are purple after staining, while Gram negative cells stain pink.<ref name="libre">{{Cite web |date=1 March 2016 |title=2.3: The Peptidoglycan Cell Wall |url=https://bio.libretexts.org/Bookshelves/Microbiology/Microbiology_(Kaiser)/Unit_1%3A_Introduction_to_Microbiology_and_Prokaryotic_Cell_Anatomy/2%3A_The_Prokaryotic_Cell_-_Bacteria/2.3%3A_The_Peptidoglycan_Cell_Wall |access-date=5 November 2023 |website=Biology LibreTexts |language=en}}</ref>

== Structure == thumb|class=skin-invert-image|Peptidoglycan. The peptidoglycan layer within the bacterial cell wall is a crystal lattice structure formed from linear chains of two alternating amino sugars, namely ''N''-acetylglucosamine (GlcNAc or NAG) and ''N''-acetylmuramic acid (MurNAc or NAM). The alternating sugars are connected by a β-(1,4)-glycosidic bond. Each MurNAc is attached to a short (4- to 5-residue) amino acid chain, containing L-alanine, D-glutamic acid, ''meso''-diaminopimelic acid, and D-alanine in the case of ''Escherichia coli'' (a gram-negative bacterium); or L-alanine, D-glutamine, L-lysine, and D-alanine with a 5-glycine interbridge between tetrapeptides in the case of ''Staphylococcus aureus'' (a gram-positive bacterium). Peptidoglycan is one of the most important sources of D-amino acids in nature.{{citation needed|date=May 2023}}

By enclosing the inner membrane, the peptidoglycan layer protects the cell from lysis caused by the turgor pressure of the cell. When the cell wall grows, it retains its shape throughout its life, so a rod shape will remain a rod shape, and a spherical shape will remain a spherical shape for life. This happens because the freshly added septal material of synthesis transforms into a hemispherical wall for the offspring cells.<ref>{{Cite journal |vauthors=Huang KC, Mukhopadhyay R, Wen B, Gitai Z, Wingreen NS |date=December 2008 |title=Cell shape and cell-wall organization in Gram-negative bacteria |journal=Proceedings of the National Academy of Sciences of the United States of America |volume=105 |issue=49 |pages=19282–19287 |bibcode=2008PNAS..10519282H |doi=10.1073/pnas.0805309105 |pmc=2592989 |pmid=19050072 |doi-access=free}}</ref>

Cross-linking between amino acids in different linear amino sugar chains occurs with the help of the enzyme DD-transpeptidase and results in a 3-dimensional structure that is strong and rigid. The specific amino acid sequence and molecular structure vary with the bacterial species.<ref>{{Cite book |title=Sherris Medical Microbiology |publisher=McGraw Hill |year=2004 |isbn=978-0-8385-8529-0 |veditors=Ryan KJ, Ray CG |edition=4th}}</ref>

The different peptidoglycan types of bacterial cell walls and their taxonomic implications have been described.<ref name="Schleifer-1972">{{Cite journal |author-link2=Otto Kandler |vauthors=Schleifer KH, Kandler O |date=December 1972 |title=Peptidoglycan types of bacterial cell walls and their taxonomic implications |journal=Bacteriological Reviews |volume=36 |issue=4 |pages=407–477 |doi=10.1128/MMBR.36.4.407-477.1972 |pmc=408328 |pmid=4568761}}</ref> Archaea (domain ''Archaea'')<ref name="Woese-1990">{{Cite journal |author-link=Carl Woese |author-link2=Otto Kandler |vauthors=Woese CR, Kandler O, Wheelis ML |date=June 1990 |title=Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya |journal=Proceedings of the National Academy of Sciences of the United States of America |volume=87 |issue=12 |pages=4576–4579 |bibcode=1990PNAS...87.4576W |doi=10.1073/pnas.87.12.4576 |pmc=54159 |pmid=2112744 |doi-access=free}}</ref> do not contain peptidoglycan (murein).<ref>{{Cite journal |author-link=Otto Kandler |vauthors=Kandler O, Hippe H |date=May 1977 |title=Lack of peptidoglycan in the cell walls of Methanosarcina barkeri |journal=Archives of Microbiology |volume=113 |issue=1–2 |pages=57–60 |bibcode=1977ArMic.113...57K |doi=10.1007/BF00428580 |pmid=889387 |s2cid=19145374}}</ref> Some Archaea contain pseudopeptidoglycan (pseudomurein, see below).<ref name="Kandler-1998">{{Cite journal |author-link=Otto Kandler |vauthors=Kandler O, König H |date=April 1998 |title=Cell wall polymers in Archaea (Archaebacteria) |journal=Cellular and Molecular Life Sciences |volume=54 |issue=4 |pages=305–308 |doi=10.1007/s000180050156 |pmc=11147200 |pmid=9614965 |s2cid=13527169}}</ref> <div class="skin-invert-image"> <gallery widths="340" heights="240"> File:Mureine.svg|The structure of peptidoglycan. NAG = ''N''-acetylglucosamine (also called GlcNAc or NAGA), NAM = ''N''-acetylmuramic acid (also called MurNAc or NAMA).

File:Gram-positive cellwall-schematic.png|Gram-positive cell wall

File:PBP catalysis.svg|Penicillin binding protein forming cross-links in newly formed bacterial cell wall. </gallery> </div> Peptidoglycan is involved in binary fission during bacterial cell reproduction. L-form bacteria and mycoplasmas, both lacking peptidoglycan cell walls, do not proliferate by binary fission, but by a budding mechanism.<ref>{{Cite journal |author-link2=Otto Kandler |vauthors=Kandler G, Kandler O |date=1954 |others=(Article in English available) |title=[Studies on morphology and multiplication of pleuropneumonia-like organisms and on bacterial L-phase, I. Light microscopy] |trans-title=Studies on morphology and multiplication of pleuropneumonia-like organisms and on bacterial L-phase, I. Light microscopy (now mycoplasmas and L-form bacteria) |journal=Archiv für Mikrobiologie |language=German |volume=21 |issue=2 |pages=178–201 |doi=10.1007/BF01816378 |pmid=14350641 |s2cid=21257985}}</ref><ref>{{Cite journal |vauthors=Leaver M, Domínguez-Cuevas P, Coxhead JM, Daniel RA, Errington J |date=February 2009 |others=[see also Erratum, 23 July 2009, Nature, vol. 460, p.538] |title=Life without a wall or division machine in Bacillus subtilis |journal=Nature |volume=457 |issue=7231 |pages=849–853 |bibcode=2009Natur.457..849L |doi=10.1038/nature07742 |pmid=19212404 |s2cid=4413852}}</ref>

In the course of early evolution, the successive development of boundaries (membranes, walls) protecting first structures of life against their environment must have been essential for the formation of the first cells (cellularisation).

The invention of rigid peptidoglycan (murein) cell walls in bacteria (domain ''Bacteria''<ref name="Woese-1990" />) was probably the prerequisite for their survival, extensive radiation and colonisation of virtually all habitats of the geosphere and hydrosphere.<ref>{{Cite book |author-link=Otto Kandler |title=Early Life on Earth. Nobel Symposium 84 |vauthors=Kandler O |publisher=Columbia U.P. |year=1994 |isbn=978-0-231-08088-0 |veditors=Bengtson S |location=New York |pages=221–270 |chapter=The early diversification of life}}</ref><ref>{{Cite book |author-link=Otto Kandler |title=Thermophiles: The keys to molecular evolution and the origin of life? |vauthors=Kandler O |publisher=Taylor and Francis Ltd. |year=1998 |isbn=978-0-203-48420-3 |veditors=Wiegel J, Adams MW |location=London |pages=19–31 |chapter=The early diversification of life and the origin of the three domains: A proposal |chapter-url=https://books.google.com/books?id=FtSzl4iastsC&q=Otto+Kandler%3A+The+early+diversification+of+life+and+the+origin+of+the+three+domains%3A+A+proposal.+In%3A+J%C3%BCrgen+Wiegel%2C+Michael+%E2%80%8EW.W.+Adams+%28Hrsg.%29%3A+Thermophiles&pg=PA19}}</ref>

== Biosynthesis == The peptidoglycan monomers are synthesized in the cytosol and are then attached to a membrane carrier bactoprenol. Bactoprenol transports peptidoglycan monomers across the cell membrane where they are inserted into the existing peptidoglycan.<ref name="The Prokaryotic Cell">{{Cite web |title=The Prokaryotic Cell: Bacteria |url=http://student.ccbcmd.edu/courses/bio141/lecguide/unit1/prostruct/cw.html |archive-url=https://web.archive.org/web/20100726231059/http://student.ccbcmd.edu/courses/bio141/lecguide/unit1/prostruct/cw.html |archive-date=26 July 2010 |access-date=1 May 2011}}</ref>

# In the first step of peptidoglycan synthesis, glutamine, which is an amino acid, donates an amino group to a sugar, fructose 6-phosphate.<ref name="White-2007" /> This reaction, catalyzed by EC 2.6.1.16 (GlmS), turns fructose 6-phosphate into glucosamine-6-phosphate.<ref name="Otten-2018">{{Cite journal |vauthors=Otten C, Brilli M, Vollmer W, Viollier PH, Salje J |date=January 2018 |title=Peptidoglycan in obligate intracellular bacteria |journal=Molecular Microbiology |volume=107 |issue=2 |pages=142–163 |doi=10.1111/mmi.13880 |pmc=5814848 |pmid=29178391 |doi-access=free}}</ref> # In step two, an acetyl group is transferred from acetyl CoA to the amino group on the glucosamine-6-phosphate creating ''N''-acetyl-glucosamine-6-phosphate.<ref name="White-2007" /> This reaction is EC 5.4.2.10, catalyzed by GlmM.<ref name="Otten-2018" /> # {{anchor|GlmU}}In step three of the synthesis process, the ''N''-acetyl-glucosamine-6-phosphate is isomerized, which will change ''N''-acetyl-glucosamine-6-phosphate to ''N''-acetyl-glucosamine-1-phosphate.<ref name="White-2007" /> This is EC 2.3.1.157, catalyzed by GlmU.<ref name="Otten-2018" /> # In step 4, the ''N''-acetyl-glucosamine-1-phosphate, which is now a monophosphate, attacks UTP. Uridine triphosphate, which is a pyrimidine nucleotide, has the ability to act as an energy source. In this particular reaction, after the monophosphate has attacked the UTP, an inorganic pyrophosphate is given off and is replaced by the monophosphate, creating UDP-N-acetylglucosamine (2,4). (When UDP is used as an energy source, it gives off an inorganic phosphate.) This initial stage, is used to create the precursor for the NAG in peptidoglycan.<ref name="White-2007" /> This is EC 2.7.7.23, also catalyzed by GlmU, which is a bifunctional enzyme.<ref name="Otten-2018" /> # In step 5, some of the UDP-N-acetylglucosamine (UDP-GlcNAc) is converted to UDP-MurNAc (UDP-N-acetylmuramic acid) by the addition of a lactyl group to the glucosamine. Also in this reaction, the C3 hydroxyl group will remove a phosphate from the alpha carbon of phosphoenolpyruvate. This creates what is called an enol derivative.<ref name="White-2007" /> EC 2.5.1.7, catalyzed by MurA.<ref name="Otten-2018" /> # In step 6, the enol is reduced to a "lactyl moiety" by NADPH in step six.<ref name="White-2007" /> EC 1.3.1.98, catalyzed by MurB.<ref name="Otten-2018" /> # In step 7, the UDP–MurNAc is converted to UDP-MurNAc pentapeptide by the addition of five amino acids, usually including the dipeptide D-alanyl-D-alanine.<ref name="White-2007" /> This is a string of three reactions: EC 6.3.2.8 by MurC, EC 6.3.2.9 by MurD, and EC 6.3.2.13 by MurE.<ref name="Otten-2018" />

Each of these reactions requires the energy source ATP.<ref name="White-2007" /> This is all referred to as Stage one.

Stage two occurs in the cytoplasmic membrane. It is in the membrane where a lipid carrier called bactoprenol carries peptidoglycan precursors through the cell membrane. # Undecaprenyl phosphate will attack the UDP-MurNAc penta, creating a PP-MurNac penta, which is now a lipid (lipid I).<ref name="White-2007" /> EC 2.7.8.13 by MraY.<ref name="Otten-2018" /> # UDP-GlcNAc is then transported to MurNAc, creating Lipid-PP-MurNAc penta-GlcNAc (lipid II), a disaccharide, also a precursor to peptidoglycan.<ref name="White-2007" /> EC 2.4.1.227 by MurG.<ref name="Otten-2018" /> # Lipid II is transported across the membrane by flippase (MurJ), a discovery made in 2014 after decades of searching.<ref name="Sham-2014">{{Cite journal |vauthors=Sham LT, Butler EK, Lebar MD, Kahne D, Bernhardt TG, Ruiz N |date=July 2014 |title=Bacterial cell wall. MurJ is the flippase of lipid-linked precursors for peptidoglycan biogenesis |journal=Science |volume=345 |issue=6193 |pages=220–222 |bibcode=2014Sci...345..220S |doi=10.1126/science.1254522 |pmc=4163187 |pmid=25013077}}</ref> Once it is there, it is added to the growing glycan chain by the enzyme peptidoglycan glycosyltransferase (GTase, EC 2.4.1.129). This reaction is known as transglycosylation. In the reaction, the hydroxyl group of the GlcNAc will attach to the MurNAc in the glycan, which will displace the lipid-PP from the glycan chain.<ref name="White-2007">{{Cite book |title=The physiology and biochemistry of prokaryotes |vauthors=White D |publisher=Oxford University Press Inc. |year=2007 |edition=3rd |location=NY}}</ref> # In a final step, the DD-transpeptidase (TPase, EC 3.4.16.4) crosslinks individual glycan chains. This protein is also known as the penicillin-binding protein. Some versions of the enzyme also performs the glycosyltransferase function, while others leave the job to a separate enzyme.<ref name="Otten-2018" /> {{clear left}}

== Pseudopeptidoglycan == {{main|Pseudopeptidoglycan}} In some archaea, i.e. members of the Methanobacteriales and in the genus ''Methanopyrus'', pseudopeptidoglycan (pseudomurein) has been found.<ref name="Kandler-1998" /> In pseudopeptidoglycan the sugar residues are β-(1,3) linked ''N''-acetylglucosamine and ''N''-acetyltalosaminuronic acid. This makes the cell walls of such archaea insensitive to lysozyme.<ref name="Madigan-2009">{{Cite book |title=Brock Biology of Microorganisms |vauthors=Madigan MT, Martinko JM, Dunlap PV, Clark DP |date=2009 |publisher=Pearson/Benjamin Cummings |edition=12th |location=San Francisco, CA}}</ref> The biosynthesis of pseudopeptidoglycan has been described.<ref>{{Cite journal |author-link2=Otto Kandler |vauthors=König H, Kandler O, Hammes W |date=January 1989 |title=Biosynthesis of pseudomurein: isolation of putative precursors from Methanobacterium thermoautotrophicum |journal=Canadian Journal of Microbiology |volume=35 |issue=1 |pages=176–181 |doi=10.1139/m89-027 |pmid=2720492}}</ref>

== Recognition by immune system == Peptidoglycan recognition is an evolutionarily conserved process.<ref name="Wolf-2018">{{Cite journal |vauthors=Wolf AJ, Underhill DM |date=April 2018 |title=Peptidoglycan recognition by the innate immune system |journal=Nature Reviews. Immunology |volume=18 |issue=4 |pages=243–254 |doi=10.1038/nri.2017.136 |pmid=29292393 |s2cid=3894187}}</ref> The overall structure is similar between bacterial species, but various modifications can increase the diversity. These include modifications of the length of sugar polymers, modifications in the sugar structures, variations in cross-linking or substitutions of amino acids (primarily at the third position).<ref name="Wolf-2018" /><ref name="Bersch-2021">{{Cite journal |display-authors=6 |vauthors=Bersch KL, DeMeester KE, Zagani R, Chen S, Wodzanowski KA, Liu S, Mashayekh S, Reinecker HC, Grimes CL |date=April 2021 |title=Bacterial Peptidoglycan Fragments Differentially Regulate Innate Immune Signaling |journal=ACS Central Science |volume=7 |issue=4 |pages=688–696 |doi=10.1021/acscentsci.1c00200 |doi-access=free|pmc=8155477 |pmid=34056099}}</ref> The aim of these modifications is to alter the properties of the cell wall, which plays a vital role in pathogenesis.<ref name="Wolf-2018" />

Peptidoglycans can be degraded by several enzymes (lysozyme, glucosaminidase, endopeptidase...<ref name="Wolf-2018" />), producing immunostimulatory fragments (sometimes called muropeptides<ref name="Bastos-2021">{{Cite journal |vauthors=Bastos PA, Wheeler R, Boneca IG |date=January 2021 |title=Uptake, recognition and responses to peptidoglycan in the mammalian host |journal=FEMS Microbiology Reviews |volume=45 |issue=1 |article-number=fuaa044 |doi=10.1093/femsre/fuaa044 |pmc=7794044 |pmid=32897324}}</ref>) that are critical for mediating host-pathogen interactions.<ref name="Bersch-2021" /> These include muramyl dipeptide (MDP), ''N''-acetylglucosamine (NAG), or γ-d-glutamyl-meso-diaminopimelic acid (iE-DAP).<ref name="Wolf-2018" /><ref name="Bastos-2021" />

Peptidoglycan from intestinal bacteria (both pathogens and commensals) crosses the intestinal barrier even under physiological conditions.<ref name="Bastos-2021" /> Mechanisms through which peptidoglycan or its fragments enter the host cells can be direct (carrier-independent) or indirect (carrier-dependent), and they are either bacteria-mediated (secretion systems, membrane vesicles) or host cell-mediated (receptor-mediated, peptide transporters).<ref name="Bastos-2021" /> Bacterial secretion systems are protein complexes used for the delivery of virulence factors across the bacterial cell envelope to the exterior environment.<ref name="Sun-2022">{{Cite journal |vauthors=Sun Q, Liu X, Li X |date=February 2022 |title=Peptidoglycan-based immunomodulation |journal=Applied Microbiology and Biotechnology |volume=106 |issue=3 |pages=981–993 |doi=10.1007/s00253-022-11795-4 |pmid=35076738 |s2cid=246276803}}</ref> Intracellular bacterial pathogens invade eukaryotic cells (which may lead to the formation of phagolysosomes and/or autophagy activation), or bacteria may be engulfed by phagocytes (macrophages, monocytes, neutrophils...). The bacteria-containing phagosome may then fuse with endosomes and lysosomes, leading to degradation of bacteria and generation of polymeric peptidoglycan fragments and muropeptides.<ref name="Bastos-2021" />

=== Receptors === Innate immune system senses intact peptidoglycan and peptidoglycan fragments using numerous PRRs (pattern recognition receptors) that are secreted, expressed intracellularly or expressed on the cell surface.<ref name="Wolf-2018" />

==== Peptidoglycan recognition proteins ==== PGLYRPs are conserved from insects to mammals.<ref name="Bastos-2021" /> Mammals produce four secreted soluble peptidoglycan recognition proteins (PGLYRP-1, PGLYRP-2, PGLYRP-3 and PGLYRP-4) that recognize muramyl pentapeptide or tetrapeptide.<ref name="Wolf-2018" /> They can also bind to LPS and other molecules by using binding sites outside of the peptidoglycan-binding groove.<ref name="Sun-2022" /> After recognition of peptidoglycan, PGLYRPs activate polyphenol oxidase (PPO) molecules, Toll, or immune deficiency (IMD) signalling pathways. That leads to production of antimicrobial peptides (AMPs).<ref name="Sun-2022" />

Each of the mammalian PGLYRPs display unique tissue expression patterns. PGLYRP-1 is mainly expressed in the granules of neutrophils and eosinophils.<ref name="Wolf-2018" /> PGLYRP-3 and 4 are expressed by several tissues such as skin, sweat glands, eyes or the intestinal tract.<ref name="Bastos-2021" /> PGLYRP-1, 3 and 4 form disulphide-linked homodimers and heterodimers essential for their bactericidal activity.<ref name="Bastos-2021" /> Their binding to bacterial cell wall peptidoglycans can induce bacterial cell death by interaction with various bacterial transcriptional regulatory proteins.<ref name="Wolf-2018" /> PGLYRPs are likely to assist in bacterial killing by cooperating with other PRRs to enhance recognition of bacteria by phagocytes.<ref name="Wolf-2018" />

PGLYRP-2 is primarily expressed by the liver and secreted into the circulation.<ref name="Wolf-2018" /> Also, its expression can be induced in skin keratinocytes, oral and intestinal epithelial cells.<ref name="Bastos-2021" /> In contrast with the other PGLYRPs, PGLYRP-2 has no direct bactericidal activity. It possesses peptidoglycan amidase activity, it hydrolyses the lactyl-amide bond between the MurNAc and the first amino acid of the stem peptide of peptidoglycan.<ref name="Wolf-2018" /><ref name="Bastos-2021" /> It is proposed, that the function of PGLYRP-2 is to prevent over-activation of the immune system and inflammation-induced tissue damage in response to NOD2 ligands (see below), as these muropeptides can no longer be recognized by NOD2 upon separation of the peptide component from MurNAc.<ref name="Bastos-2021" /> Growing evidence suggests that peptidoglycan recognition protein family members play a dominant role in the tolerance of intestinal epithelial cells toward the commensal microbiota.<ref name="Sun-2022" /><ref>{{Cite journal |vauthors=Liang Y, Yang L, Wang Y, Tang T, Liu F, Zhang F |date=December 2022 |title=Peptidoglycan recognition protein SC (PGRP-SC) shapes gut microbiota richness, diversity and composition by modulating immunity in the house fly Musca domestica |journal=Insect Molecular Biology |volume=32 |issue=2 |pages=200–212 |doi=10.1111/imb.12824 |pmid=36522831 |s2cid=254807823}}</ref> It has been demonstrated that expression of PGLYRP-2 and 4 can influence the composition of the intestinal microbiota.<ref name="Sun-2022" />

Recently, it has been discovered, that PGLYRPs (and also NOD-like receptors and peptidoglycan transporters) are highly expressed in the developing mouse brain.<ref name="Gonzalez-Santana-2020">{{Cite journal |vauthors=Gonzalez-Santana A, Diaz Heijtz R |date=August 2020 |title=Bacterial Peptidoglycans from Microbiota in Neurodevelopment and Behavior |url=https://hal.archives-ouvertes.fr/hal-03492013/file/S1471491420301325.pdf |journal=Trends in Molecular Medicine |volume=26 |issue=8 |pages=729–743 |doi=10.1016/j.molmed.2020.05.003 |pmid=32507655 |s2cid=219539658}}</ref> PGLYRP-2 and is highly expressed in neurons of several brain regions including the prefrontal cortex, hippocampus, and cerebellum, thus indicating potential direct effects of peptidoglycan on neurons. PGLYRP-2 is highly expressed also in the cerebral cortex of young children, but not in most adult cortical tissues. PGLYRP-1 is also expressed in the brain and continues to be expressed into adulthood.<ref name="Gonzalez-Santana-2020" />

==== NOD-like receptors ==== Probably the most well-known receptors of peptidoglycan are the NOD-like receptors (NLRs), mainly NOD1 and NOD2. The NOD1 receptor is activated after iE-DAP (γ-d-glutamyl-meso-diaminopimelic acid) binding, while NOD2 recognizes MDP (muramyl dipeptide), by their LRR domains.<ref name="Sun-2022" /> Activation leads to self-oligomerization, resulting in activation of two signalling cascades. One triggers activation of NF-κB (through RIP2, TAK1 and IKK<ref name="Murphy-2017">{{Cite book |title=Janeway's immunobiology |vauthors=Murphy K, Weaver C, Janeway C |publisher=Garland Science |year=2017 |isbn=978-0-8153-4505-3 |edition=9th |location=New York |pages=45, 96–98 |oclc=933586700}}</ref>), second leads to MAPK signalling cascade. Activation of these pathways induces production of inflammatory cytokines and chemokines.<ref name="Wolf-2018" />

NOD1 is expressed by diverse cell types, including myeloid phagocytes, epithelial cells<ref name="Wolf-2018" /> and neurons.<ref name="Gonzalez-Santana-2020" /> NOD2 is expressed in monocytes and macrophages, epithelial intestinal cells, Paneth cells, dendritic cells, osteoblasts, keratinocytes and other epithelial cell types.<ref name="Bastos-2021" /> As cytosolic sensors, NOD1 and NOD2 must either detect bacteria that enter the cytosol, or peptidoglycan must be degraded to generate fragments that must be transported into the cytosol for these sensors to function.<ref name="Wolf-2018" />

Recently, it was demonstrated that NLRP3 is activated by peptidoglycan, through a mechanism that is independent of NOD1 and NOD2.<ref name="Bastos-2021" /> In macrophages, N-acetylglucosamine generated by peptidoglycan degradation was found to inhibit hexokinase activity and induce its release from the mitochondrial membrane. It promotes NLRP3 inflammasome activation through a mechanism triggered by increased mitochondrial membrane permeability.<ref name="Bastos-2021" />

NLRP1 is also considered as a cytoplasmic sensor of peptidoglycan. It can sense MDP and promote IL-1 secretion through binding NOD2.<ref name="Sun-2022" /><ref name="Bersch-2021" />

==== C-type lectin receptors (CLRs) ==== C-type lectins are a diverse superfamily of mainly Ca2+-dependent proteins that bind a variety of carbohydrates (including the glycan skeleton of peptidoglycan), and function as innate immune receptors.<ref name="Bastos-2021" /> CLR proteins that bind to peptidoglycan include mannose binding lectin (MBL), ficolins, Reg3A (regeneration gene family protein 3A), and PTCLec1.<ref name="Sun-2022" /> In mammals, they initiate the lectin-pathway of the complement cascade.<ref name="Bastos-2021" />

==== Toll-like receptors ==== The role of toll-like receptors (TLRs) in direct recognition of peptidoglycan is controversial.<ref name="Wolf-2018" /> In some studies, has been reported that peptidoglycan is sensed by TLR2.<ref>{{Cite journal |vauthors=Yoshimura A, Lien E, Ingalls RR, Tuomanen E, Dziarski R, Golenbock D |date=July 1999 |title=Cutting edge: recognition of Gram-positive bacterial cell wall components by the innate immune system occurs via Toll-like receptor 2 |journal=Journal of Immunology |volume=163 |issue=1 |pages=1–5 |doi=10.4049/jimmunol.163.1.1 |pmid=10384090 |s2cid=23630870 |doi-access=free}}</ref> But this TLR2-inducing activity could be due to cell wall lipoproteins and lipoteichoic acids that commonly co-purify with peptidoglycan. Also variation in peptidoglycan structure in bacteria from species to species may contribute to the differing results on this topic.<ref name="Wolf-2018" /><ref name="Bastos-2021" />

== As vaccine or adjuvant == Peptidoglycan is immunologically active, which can stimulate immune cells to increase the expression of cytokines and enhance antibody-dependent specific response when combined with vaccine or as adjuvant alone.<ref name="Sun-2022" /> MDP, which is the basic unit of peptidoglycan, was initially used as the active component of Freund's adjuvant.<ref name="Sun-2022" /> Peptidoglycan from ''Staphylococcus aureus'' was used as a vaccine to protect mice, showing that after vaccine injection for 40 weeks, the mice survived from ''S. aureus'' challenge at an increased lethal dose.<ref>{{Cite journal |vauthors=Capparelli R, Nocerino N, Medaglia C, Blaiotta G, Bonelli P, Iannelli D |date=2011-12-01 |title=The Staphylococcus aureus peptidoglycan protects mice against the pathogen and eradicates experimentally induced infection |journal=PLOS ONE |volume=6 |issue=12 |article-number=e28377 |bibcode=2011PLoSO...628377C |doi=10.1371/journal.pone.0028377 |pmc=3228750 |pmid=22145040 |doi-access=free |veditors=Cardona PJ}}</ref>

== Inhibition and degradation == Some antibacterial drugs such as penicillin interfere with the production of peptidoglycan by binding to bacterial enzymes known as penicillin-binding proteins or DD-transpeptidases.<ref name="Salton-1996" /> Penicillin-binding proteins form the bonds between oligopeptide crosslinks in peptidoglycan. For a bacterial cell to reproduce through binary fission, more than a million peptidoglycan subunits (NAM-NAG+oligopeptide) must be attached to existing subunits.<ref>{{Cite book |last=Bauman R |title=Microbiology with Diseases by Taxonomy |publisher=Benjamin Cummings |year=2007 |isbn=978-0-8053-7679-1 |edition=2nd}}</ref> Mutations in genes coding for transpeptidases that lead to reduced interactions with an antibiotic are a significant source of emerging antibiotic resistance.<ref>{{Cite journal |vauthors=Spratt BG |date=April 1994 |title=Resistance to antibiotics mediated by target alterations |journal=Science |volume=264 |issue=5157 |pages=388–393 |bibcode=1994Sci...264..388S |doi=10.1126/science.8153626 |pmid=8153626 |s2cid=30578841}}</ref> Since peptidoglycan is also lacking in L-form bacteria and in mycoplasmas, both are resistant against penicillin.

Other steps of peptidoglycan synthesis can also be targeted. The topical antibiotic bacitracin targets the utilization of C55-isoprenyl pyrophosphate. Lantibiotics, which include the food preservative nisin, attack lipid II.<ref>{{Cite journal |vauthors=Sarkar P, Yarlagadda V, Ghosh C, Haldar J |date=March 2017 |title=A review on cell wall synthesis inhibitors with an emphasis on glycopeptide antibiotics |journal=MedChemComm |volume=8 |issue=3 |pages=516–533 |doi=10.1039/c6md00585c |pmc=6072328 |pmid=30108769}}</ref>

Lysozyme, which is found in tears and constitutes part of the body's innate immune system exerts its antibacterial effect by breaking the β-(1,4)-glycosidic bonds in peptidoglycan (see above). Lysozyme is more effective in acting against gram-positive bacteria, in which the peptidoglycan cell wall is exposed, than against gram-negative bacteria, which have an outer layer of LPS covering the peptidoglycan layer.<ref name="Murphy-2017" /> Several bacterial peptidoglycan modifications can result in resistance to degradation by lysozyme. Susceptibility of bacteria to degradation is also considerably affected by exposure to antibiotics. Exposed bacteria synthesize peptidoglycan that contains shorter sugar chains that are poorly crosslinked and this peptidoglycan is then more easily degraded by lysozyme.<ref name="Sun-2022" />

== Peptidoglycan as a danger signal among bacteria == A 2025 study reported that peptidoglycan fragments released from lysed bacterial cells can function as a general danger signal among diverse bacterial species.<ref name=":0">{{Cite journal|title=Bacteria use exogenous peptidoglycan as a danger signal to trigger biofilm formation|url=https://www.nature.com/articles/s41564-024-01886-5|journal=Nature Microbiology|date=January 2025|issn=2058-5276|pmc=11726461|pmid=39753671|pages=144–157|volume=10|issue=1|doi=10.1038/s41564-024-01886-5|language=en|first=Sanika|last=Vaidya|first2=Dibya|last2=Saha|first3=Daniel K. H.|last3=Rode|first4=Gabriel|last4=Torrens|first5=Mads F.|last5=Hansen|first6=Praveen K.|last6=Singh|first7=Eric|last7=Jelli|first8=Kazuki|last8=Nosho|first9=Hannah|last9=Jeckel|first10=Stephan|last10=Göttig|first11=Felipe|last11=Cava|first12=Knut|last12=Drescher}}</ref> Exposure to exogenous peptidoglycan was shown to rapidly induce the formation of three‑dimensional biofilms in ''Vibrio cholerae'', ''Pseudomonas aeruginosa, Staphylococcus aureus'', ''Acinetobacter baumannii'' and ''Enterococcus faecalis''. Even brief exposure was sufficient to trigger a regulated response leading to increased production of biofilm matrix components. In ''V. cholerae'', peptidoglycan exposure upregulated several genes involved in matrix synthesis, including the ''vps''‑I and ''vps''‑II gene clusters that contribute to biofilm structure. The mechanism by which bacteria sense extracellular peptidoglycan fragments remains unknown.<ref name=":0" />

== See also == * Undecaprenyl-diphosphatase

== References == {{Reflist}}

== External links == * [https://www.ncbi.nlm.nih.gov/books/NBK8477/figure/A298/ Diagrammatic representation of peptidoglycan structures.] * [http://pubs.acs.org/doi/abs/10.1021/bi4010446 Structure of MurNAc 6-Phosphate Hydrolase (MurQ) from Haemophilus influenzae with a Bound Inhibitor.] {{Mucoproteins}} {{Bacteria}}

Category:Membrane biology Category:Glycobiology