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Haobo Jiang - One of the best experts on this subject based on the ideXlab platform.

  • The three-dimensional structure and recognition mechanism of Manduca sexta peptidoglycan recognition protein-1.
    Insect biochemistry and molecular biology, 2019
    Co-Authors: Xiaolong Cao, Yang Wang, Geert-jan Boons, Junpeng Deng, Haobo Jiang
    Abstract:

    Peptidoglycan recognition proteins (PGRPs) recognize bacteria through their unique cell wall constituent, Peptidoglycans (PGs). PGRPs are conserved from insects to mammals and all function in antibacterial defense. In the tobacco hornworm Manduca sexta, PGRP1 and microbe binding protein (MBP) interact with PGs and hemolymph protease-14 precursor (proHP14) to yield active HP14. HP14 triggers a serine protease network that produces active phenoloxidase (PO), Spatzle, and other cytokines to stimulate immune responses. PGRP1 binds preferentially to diaminopimelic acid (DAP)-PGs of Gram-negative bacteria and Gram-positive Bacillus and Clostridium species than Lys-PGs of other Gram-positive bacteria. In this study, we synthesized DAP- and Lys-muramyl pentapeptide (MPP) and monitored their associations with M. sexta PGRP1 by surface plasmon resonance. The Kd values (0.57 μM for DAP-MPP and 45.6 μM for Lys-MPP) agree with the differential recognition of DAP- and Lys-PGs. To reveal its structural basis, we produced the PGRP1 in insect cells and determined its structure at a resolution of 2.1 A. The protein adopts a fold similar to those from other PGRPs with a classical L-shaped PG-binding groove. A unique loop lining the shallow groove suggests a different ligand-binding mechanism. In summary, this study provided new insights into the PG recognition by PGRPs, a critical first step that initiates the serine protease cascade.

  • a short type peptidoglycan recognition protein from the silkworm expression characterization and involvement in the prophenoloxidase activation pathway
    Developmental and Comparative Immunology, 2014
    Co-Authors: Kangkang Chen, Chen Liu, Haobo Jiang
    Abstract:

    Recognition of invading microbes as non-self is the first step of immune responses. In insects, peptidoglycan recognition proteins (PGRPs) detect Peptidoglycans (PGs) of bacterial cell wall, leading to the activation of defense responses. Twelve PGRPs have been identified in the silkworm, Bombyx mori, through bioinformatics analysis. However, their biochemical functions are mostly uncharacterized. In this study, we found PGRP-S5 transcript levels were up-regulated in fat body and midgut after bacterial infection. Using recombinant protein isolated from Escherichia coli, we showed that PGRP-S5 binds to PGs from certain bacterial strains and induces bacteria agglutination. Enzyme activity assay confirmed PGRP-S5 is an amidase; we also showed it is an antibacterial protein effective against both Gram-positive and -negative bacteria. Additionally, we demonstrated that specific recognition of PGs by PGRP-S5 is involved in the prophenoloxidase activation pathway. Together, these data suggest the silkworm PGRP-S5 functions as a pattern recognition receptor for the prophenoloxidase pathway initiation and as an effecter to inhibit bacterial growth as well. We finally discussed possible roles of PGRP-S5 as a receptor for antimicrobial peptide gene induction and as an immune modulator in the midgut.

  • involvement of manduca sexta peptidoglycan recognition protein 1 in the recognition of bacteria and activation of prophenoloxidase system
    Insect Biochemistry and Molecular Biology, 2010
    Co-Authors: Niranji Sumathipala, Haobo Jiang
    Abstract:

    Although the importance of peptidoglycan recognition proteins (PGRPs) in detecting bacteria and promoting immunity is well recognized in Drosophila melanogaster and other insect species, such a role has not yet been experimentally established for PGRPs in the tobacco hornworm, Manduca sexta. In this study, we purified M. sexta PGRP1 from the baculovirus-insect cell expression system, tested its association with Peptidoglycans and intact bacteria, and explored its possible link with the prophenoloxidase activation system in larval hemolymph. Sequence comparison suggested that PGRP1 is not an amidase and lacks residues for interacting with the carboxyl group of meso-diaminopimelic acid-Peptidoglycans (DAP-PGs). M. sexta PGRP1 gene was constitutively expressed at a low level in fat body, and the mRNA concentration became much higher after an injection of Escherichia coli. Consistently, the protein concentration in larval plasma increased in a time-dependent manner after the immune challenge. Purified recombinant PGRP1 specifically bound to soluble DAP-PG of E. coli but not to soluble Lys-type PG of Staphylococcus aureus. In addition, this recognition protein completely bound to insoluble PGs from Micrococcus luteus, Bacillus megaterium and Bacillus subtilis, whereas its association with the bacterial cells was low even though their Peptidoglycans are exposed on the cell surface. After PGRP1 had been added to plasma of naive larvae in the absence of microbial elicitor, there was a concentration-dependent increase in prophenoloxidase activation. Phenoloxidase activity, as usual, increased after the plasma was incubated with peptidoglyans or bacterial cells. These increases became more prominent when insoluble M. luteus or B. megaterium PG or soluble E. coli PG and PGRP1 were both present. Statistic analysis suggested a synergistic effect caused by interaction between PGRP1 and these PGs. Taken together, these results indicated that PGRP1 is a member of the M. sexta prophenoloxidase activation system, which recognizes Peptidoglycans from certain bacteria and initiates the host defense response. The unexplained difference between the purified PGs and intact bacteria clearly reflects our general lack of understanding of PGRP1-mediated recognition and how it leads to proPO activation.

Roy A. Mariuzza - One of the best experts on this subject based on the ideXlab platform.

  • activity augmentation of amphioxus peptidoglycan recognition protein bbtpgrp3 via fusion with a chitin binding domain
    PLOS ONE, 2015
    Co-Authors: Wenjie Wang, Roy A. Mariuzza, Wang Cheng, Ming Luo, Qingyu Yan, Dongdong Cao, Shengfeng Huang, Yuxing Chen, Congzhao Zhou
    Abstract:

    Peptidoglycan recognition proteins (PGRPs), which have been identified in most animals, are pattern recognition molecules that involve antimicrobial defense. Resulting from extraordinary expansion of innate immune genes, the amphioxus encodes many PGRPs of diverse functions. For instance, three isoforms of PGRP encoded by Branchiostoma belcheri tsingtauense, termed BbtPGRP1~3, are fused with a chitin binding domain (CBD) at the N-terminus. Here we report the 2.7 A crystal structure of BbtPGRP3, revealing an overall structure of an N-terminal hevein-like CBD followed by a catalytic PGRP domain. Activity assays combined with site-directed mutagenesis indicated that the individual PGRP domain exhibits amidase activity towards both DAP-type and Lys-type Peptidoglycans (PGNs), the former of which is favored. The N-terminal CBD not only has the chitin-binding activity, but also enables BbtPGRP3 to gain a five-fold increase of amidase activity towards the Lys-type PGNs, leading to a significantly broadened substrate spectrum. Together, we propose that modular evolution via domain shuffling combined with gene horizontal transfer makes BbtPGRP1~3 novel PGRPs of augmented catalytic activity and broad recognition spectrum.

  • crystal structure of human peptidoglycan recognition protein iα bound to a muramyl pentapeptide from gram positive bacteria
    Protein Science, 2006
    Co-Authors: Rongjin Guan, Geert-jan Boons, Patrick H Brown, Chittoor P Swaminathan, Abhijit Roychowdhury, Roy A. Mariuzza
    Abstract:

    Peptidoglycan recognition proteins (PGRPs) are pattern recognition receptors of the innate immune system that bind bacterial Peptidoglycans (PGNs). We determined the crystal structure, to 2.1 A resolution, of the C-terminal PGN-binding domain of human PGRP-Iα in complex with a muramyl pentapeptide (MPP) from Gram-positive bacteria containing a complete peptide stem (L-Ala-D-isoGln-L-Lys-D-Ala-D-Ala). The structure reveals important features not observed previously in the complex between PGRP-Iα and a muramyl tripeptide lacking D-Ala at stem positions 4 and 5. Most notable are ligand-induced structural rearrangements in the PGN-binding site that are essential for entry of the C-terminal portion of the peptide stem and for locking MPP in the binding groove. We propose that similar structural rearrangements to accommodate the PGN stem likely characterize many PGRPs, both mammalian and insect.

  • Crystal structure of a peptidoglycan recognition protein (PGRP) in complex with a muramyl tripeptide from Gram-positive bacteria.
    Journal of endotoxin research, 2005
    Co-Authors: Rongjin Guan, Geert-jan Boons, Abhijit Roychowdury, Brian Ember, Sanjay Kumar, Roy A. Mariuzza
    Abstract:

    Peptidoglycan recognition proteins (PGRPs) are pattern recognition receptors of the innate immune system that bind, and in some cases hydrolyse, bacterial Peptidoglycans (PGNs). We determined the crystal structure of the C-terminal PGN-binding domain of human PGRP-Iα in complex with a muramyl tripeptide representing the conserved core of lysine-type PGNs. The peptide stem of the ligand is buried at the deep end of a long binding groove, with N-acetylmuramic acid situated in the middle of the groove, whose shallow end could accommodate N-acetylglucosamine. Both peptide and glycan moieties are essential for binding by PGRPs. Conservation of key PGN-contacting residues indicates that all PGRPs employ this basic PGN-binding mode. The structure identifies variable residues that likely mediate discrimination between lysine- and diaminopimelic acid-type PGNs. In addition, we propose a mechanism for PGN hydrolysis by Zn2+-containing catalytic PGRPs.

  • crystal structure of the c terminal peptidoglycan binding domain of human peptidoglycan recognition protein ialpha
    Journal of Biological Chemistry, 2004
    Co-Authors: Rongjin Guan, Emilio L Malchiodi, Qian Wang, Peter Schuck, Roy A. Mariuzza
    Abstract:

    Abstract Peptidoglycan recognition proteins (PGRPs) are pattern recognition receptors of the innate immune system that bind, and in some cases hydrolyze, Peptidoglycans (PGNs) on bacterial cell walls. These molecules, which are highly conserved from insects to mammals, participate in host defense against both Gram-positive and Gram-negative bacteria. We report the crystal structure of the C-terminal PGN-binding domain of human PGRP-Iα in two oligomeric states, monomer and dimer, to resolutions of 2.80 and 1.65 A, respectively. In contrast to PGRPs with PGN-lytic amidase activity, no zinc ion is present in the PGN-binding site of human PGRP-Iα. The structure reveals that PGRPs exhibit extensive topological variability in a large hydrophobic groove, located opposite the PGN-binding site, which may recognize host effector proteins or microbial ligands other than PGN. We also show that full-length PGRP-Iα comprises two tandem PGN-binding domains. These domains differ at most potential PGN-contacting positions, implying different fine specificities. Dimerization of PGRP-Iα, which occurs through three-dimensional domain swapping, is mediated by specific binding of sodium ions to a flexible hinge loop, stabilizing the conformation found in the dimer. We further demonstrate sodium-dependent dimerization of PGRP-Iα in solution, suggesting a possible mechanism for modulating PGRP activity through the formation of multivalent adducts.

Thomas G Bernhardt - One of the best experts on this subject based on the ideXlab platform.

  • ftsw is a peptidoglycan polymerase that is functional only in complex with its cognate penicillin binding protein
    Nature microbiology, 2019
    Co-Authors: Atsushi Taguchi, Daniel Kahne, Andrew C Kruse, Thomas G Bernhardt, Michael Welsh, Lindsey S Marmont, Wonsik Lee, Megan Sjodt
    Abstract:

    The peptidoglycan cell wall is essential for the survival and morphogenesis of bacteria1. For decades, it was thought that only class A penicillin-binding proteins (PBPs) and related enzymes effected peptidoglycan synthesis. Recently, it was shown that RodA-a member of the unrelated SEDS protein family-also acts as a peptidoglycan polymerase2-4. Not all bacteria require RodA for growth; however, its homologue, FtsW, is a core member of the divisome complex that appears to be universally essential for septal cell wall assembly5,6. FtsW was previously proposed to translocate the peptidoglycan precursor lipid II across the cytoplasmic membrane7,8. Here, we report that purified FtsW polymerizes lipid II into peptidoglycan, but show that its polymerase activity requires complex formation with its partner class B PBP. We further demonstrate that the polymerase activity of FtsW is required for its function in vivo. Thus, our findings establish FtsW as a peptidoglycan polymerase that works with its cognate class B PBP to produce septal peptidoglycan during cell division.

  • ftsw is a peptidoglycan polymerase that is activated by its cognate penicillin binding protein
    bioRxiv, 2018
    Co-Authors: Atsushi Taguchi, Daniel Kahne, Thomas G Bernhardt, Michael Welsh, Lindsey S Marmont, Wonsik Lee, Suzanne Walker
    Abstract:

    The peptidoglycan cell wall is essential for the survival and shape maintenance of bacteria. For decades it was thought that only penicillin-binding proteins (PBPs) effected peptidoglycan synthesis. Recently, it was shown that RodA, a member of the Rod complex involved in side wall peptidoglycan synthesis, acts as a peptidoglycan polymerase. RodA is absent or dispensable in many bacteria that contain a cell wall; however, all of these bacteria have a RodA homologue, FtsW, which is a core member of the divisome complex that is essential for septal cell wall assembly. FtsW was previously proposed flip the peptidoglycan precursor Lipid II to the peripasm, but we report here that FtsW polymerizes Lipid II. We show that FtsW polymerase activity depends on the presence of the class B PBP (bPBP) that it recruits to the septum. We also demonstrate that the polymerase activity of FtsW is required for its function in vivo. Our findings establish FtsW as a peptidoglycan polymerase that works with its cognate bPBP to produce septal peptidoglycan during cell division.

  • seds proteins are a widespread family of bacterial cell wall polymerases
    Nature, 2016
    Co-Authors: Alexander J Meeske, Eammon P Riley, William P Robins, Tsuyoshi Uehara, John J Mekalanos, Daniel Kahne, Suzanne Walker, Andrew C Kruse, Thomas G Bernhardt, David Z Rudner
    Abstract:

    Elongation of rod-shaped bacteria is mediated by a dynamic peptidoglycan-synthetizing machinery called the Rod complex. Here we report that, in Bacillus subtilis, this complex is functional in the absence of all known peptidoglycan polymerases. Cells lacking these enzymes survive by inducing an envelope stress response that increases the expression of RodA, a widely conserved core component of the Rod complex. RodA is a member of the SEDS (shape, elongation, division and sporulation) family of proteins, which have essential but ill-defined roles in cell wall biogenesis during growth, division and sporulation. Our genetic and biochemical analyses indicate that SEDS proteins constitute a family of peptidoglycan polymerases. Thus, B. subtilis and probably most bacteria use two distinct classes of polymerase to synthesize their exoskeleton. Our findings indicate that SEDS family proteins are core cell wall synthases of the cell elongation and division machinery, and represent attractive targets for antibiotic development. SEDS proteins are core peptidoglycan polymerases involved in bacterial cell wall elongation and division. It has been generally accepted that the cell wall Peptidoglycans of the bacterial exoskeleton are synthesized by penicillin binding proteins (PBPs) known as class A PBPs. Now, using genetic manipulation, phylogenetic analysis and functional experiments in Bacillus subtilis, David Rudner and colleagues have identified SEDS family proteins as the main peptidoglycan polymerases more broadly conserved than class A PBPs. Specifically in B. subtilis, they show that the SEDS protein RodA, a widely conserved component of the Rod complex involved in elongation of rod-shaped bacteria, acts with class B PBPs as the core cell wall synthase of the cell elongation and division machinery. The authors conclude that B. subtilis and probably most bacteria use two distinct classes of polymerases to synthesize their exoskeleton. This work also suggests that SEDS family proteins should be attractive targets for antibiotic development.

Suzanne Walker - One of the best experts on this subject based on the ideXlab platform.

  • uncovering the activities biological roles and regulation of bacterial cell wall hydrolases and tailoring enzymes
    Journal of Biological Chemistry, 2020
    Co-Authors: Julia E Page, Suzanne Walker
    Abstract:

    Bacteria account for 1000-fold more biomass than humans. They vary widely in shape and size. The morphological diversity of bacteria is due largely to the different peptidoglycan-based cell wall structures that encase bacterial cells. Although the basic structure of peptidoglycan is highly conserved, consisting of long glycan strands that are cross-linked by short peptide chains, the mature cell wall is chemically diverse. Peptidoglycan hydrolases and cell wall-tailoring enzymes that regulate glycan strand length, the degree of cross-linking, and the addition of other modifications to peptidoglycan are central in determining the final architecture of the bacterial cell wall. Historically, it has been difficult to biochemically characterize these enzymes that act on peptidoglycan because suitable peptidoglycan substrates were inaccessible. In this review, we discuss fundamental aspects of bacterial cell wall synthesis, describe the regulation and diverse biochemical and functional activities of peptidoglycan hydrolases, and highlight recently developed methods to make and label defined peptidoglycan substrates. We also review how access to these substrates has now enabled biochemical studies that deepen our understanding of how bacterial cell wall enzymes cooperate to build a mature cell wall. Such improved understanding is critical to the development of new antibiotics that disrupt cell wall biogenesis, a process essential to the survival of bacteria.

  • staphylococcus aureus cell growth and division are regulated by an amidase that trims peptides from uncrosslinked peptidoglycan
    Nature microbiology, 2020
    Co-Authors: Kaitlin Schaefer, Daniel Kahne, Mariana G Pinho, Ace George Santiago, Pedro B. Fernandes, Kathryn A Coe, Suzanne Walker
    Abstract:

    Bacteria are protected by a polymer of peptidoglycan that serves as an exoskeleton1. In Staphylococcus aureus, the peptidoglycan assembly enzymes relocate during the cell cycle from the periphery, where they are active during growth, to the division site where they build the partition between daughter cells2-4. But how peptidoglycan synthesis is regulated throughout the cell cycle is poorly understood5,6. Here, we used a transposon screen to identify a membrane protein complex that spatially regulates S. aureus peptidoglycan synthesis. This complex consists of an amidase that removes stem peptides from uncrosslinked peptidoglycan and a partner protein that controls its activity. Amidases typically hydrolyse crosslinked peptidoglycan between daughter cells so that they can separate7. However, this amidase controls cell growth. In its absence, peptidoglycan synthesis becomes spatially dysregulated, which causes cells to grow so large that cell division is defective. We show that the cell growth and division defects due to loss of this amidase can be mitigated by attenuating the polymerase activity of the major S. aureus peptidoglycan synthase. Our findings lead to a model wherein the amidase complex regulates the density of peptidoglycan assembly sites to control peptidoglycan synthase activity at a given subcellular location. Removal of stem peptides from peptidoglycan at the cell periphery promotes peptidoglycan synthase relocation to midcell during cell division. This mechanism ensures that cell expansion is properly coordinated with cell division.

  • the cell cycle in staphylococcus aureus is regulated by an amidase that controls peptidoglycan synthesis
    bioRxiv, 2019
    Co-Authors: Kaitlin Schaefer, Daniel Kahne, Mariana G Pinho, Ace George Santiago, Pedro B. Fernandes, Kathryn A Coe, Suzanne Walker
    Abstract:

    Abstract Bacteria are protected by a polymer of peptidoglycan that serves as an exoskeleton. In Staphylococcus aureus, the enzymes that assemble peptidoglycan move during the cell cycle from the periphery, where they are active during growth, to the division site where they build the partition between daughter cells. But how peptidoglycan synthesis is regulated throughout the cell cycle is not understood. Here we identify a membrane protein complex that spatially regulates S. aureus peptidoglycan synthesis. This complex consists of an amidase that removes peptide chains from uncrosslinked peptidoglycan and a partner protein that controls its activity. Typical amidases act after cell division to hydrolyze peptidoglycan between daughter cells so they can separate. However, we show that this amidase controls cell growth. In its absence, excess peptidoglycan synthesis occurs at the cell periphery, causing cells to grow so large that cell division is defective. We show that cell growth and division defects due to loss of this amidase can be mitigated by attenuating the polymerase activity of the major S. aureus peptidoglycan synthase. Our findings lead to a model wherein the amidase complex regulates the density of peptidoglycan assembly sites to control peptidoglycan synthase activity at a given cellular location. Removal of peptide chains from peptidoglycan at the cell periphery promotes synthase movement to midcell during cell division. This mechanism ensures that cell expansion is properly coordinated with cell division.

  • ftsw is a peptidoglycan polymerase that is activated by its cognate penicillin binding protein
    bioRxiv, 2018
    Co-Authors: Atsushi Taguchi, Daniel Kahne, Thomas G Bernhardt, Michael Welsh, Lindsey S Marmont, Wonsik Lee, Suzanne Walker
    Abstract:

    The peptidoglycan cell wall is essential for the survival and shape maintenance of bacteria. For decades it was thought that only penicillin-binding proteins (PBPs) effected peptidoglycan synthesis. Recently, it was shown that RodA, a member of the Rod complex involved in side wall peptidoglycan synthesis, acts as a peptidoglycan polymerase. RodA is absent or dispensable in many bacteria that contain a cell wall; however, all of these bacteria have a RodA homologue, FtsW, which is a core member of the divisome complex that is essential for septal cell wall assembly. FtsW was previously proposed flip the peptidoglycan precursor Lipid II to the peripasm, but we report here that FtsW polymerizes Lipid II. We show that FtsW polymerase activity depends on the presence of the class B PBP (bPBP) that it recruits to the septum. We also demonstrate that the polymerase activity of FtsW is required for its function in vivo. Our findings establish FtsW as a peptidoglycan polymerase that works with its cognate bPBP to produce septal peptidoglycan during cell division.

  • seds proteins are a widespread family of bacterial cell wall polymerases
    Nature, 2016
    Co-Authors: Alexander J Meeske, Eammon P Riley, William P Robins, Tsuyoshi Uehara, John J Mekalanos, Daniel Kahne, Suzanne Walker, Andrew C Kruse, Thomas G Bernhardt, David Z Rudner
    Abstract:

    Elongation of rod-shaped bacteria is mediated by a dynamic peptidoglycan-synthetizing machinery called the Rod complex. Here we report that, in Bacillus subtilis, this complex is functional in the absence of all known peptidoglycan polymerases. Cells lacking these enzymes survive by inducing an envelope stress response that increases the expression of RodA, a widely conserved core component of the Rod complex. RodA is a member of the SEDS (shape, elongation, division and sporulation) family of proteins, which have essential but ill-defined roles in cell wall biogenesis during growth, division and sporulation. Our genetic and biochemical analyses indicate that SEDS proteins constitute a family of peptidoglycan polymerases. Thus, B. subtilis and probably most bacteria use two distinct classes of polymerase to synthesize their exoskeleton. Our findings indicate that SEDS family proteins are core cell wall synthases of the cell elongation and division machinery, and represent attractive targets for antibiotic development. SEDS proteins are core peptidoglycan polymerases involved in bacterial cell wall elongation and division. It has been generally accepted that the cell wall Peptidoglycans of the bacterial exoskeleton are synthesized by penicillin binding proteins (PBPs) known as class A PBPs. Now, using genetic manipulation, phylogenetic analysis and functional experiments in Bacillus subtilis, David Rudner and colleagues have identified SEDS family proteins as the main peptidoglycan polymerases more broadly conserved than class A PBPs. Specifically in B. subtilis, they show that the SEDS protein RodA, a widely conserved component of the Rod complex involved in elongation of rod-shaped bacteria, acts with class B PBPs as the core cell wall synthase of the cell elongation and division machinery. The authors conclude that B. subtilis and probably most bacteria use two distinct classes of polymerases to synthesize their exoskeleton. This work also suggests that SEDS family proteins should be attractive targets for antibiotic development.

Alyssa Panitch - One of the best experts on this subject based on the ideXlab platform.

  • collagen binding Peptidoglycans inhibit mmp mediated collagen degradation and reduce dermal scarring
    PLOS ONE, 2011
    Co-Authors: Kate Stuart, John E Paderi, Lynetta J Freeman, Paul W. Snyder, Alyssa Panitch
    Abstract:

    Scarring of the skin is a large unmet clinical problem that is of high patient concern and impact. Wound healing is complex and involves numerous pathways that are highly orchestrated, leaving the skin sealed, but with abnormal organization and composition of tissue components, namely collagen and proteoglycans, that are then remodeled over time. To improve healing and reduce or eliminate scarring, more rapid restoration of healthy tissue composition and organization offers a unique approach for development of new therapeutics. A synthetic collagen-binding peptidoglycan has been developed that inhibits matrix metalloproteinase-1 and 13 (MMP-1 and MMP-13) mediated collagen degradation. We investigated the synthetic peptidoglycan in a rat incisional model in which a single dose was delivered in a hyaluronic acid (HA) vehicle at the time of surgery prior to wound closure. The peptidoglycan treatment resulted in a significant reduction in scar tissue at 21 days as measured by histology and visual analysis. Improved collagen architecture of the treated wounds was demonstrated by increased tensile strength and transmission electron microscopy (TEM) analysis of collagen fibril diameters compared to untreated and HA controls. The peptidoglycan's mechanism of action includes masking existing collagen and inhibiting MMP-mediated collagen degradation while modulating collagen organization. The peptidoglycan can be synthesized at low cost with unique design control, and together with demonstrated preclinical efficacy in reducing scarring, warrants further investigation for dermal wound healing.

  • the inhibition of platelet adhesion and activation on collagen during balloon angioplasty by collagen binding Peptidoglycans
    Biomaterials, 2011
    Co-Authors: John E Paderi, Kate Stuart, Michael Sturek, Kinam Park, Alyssa Panitch
    Abstract:

    Collagen is a potent stimulator for platelet adhesion, activation, and thrombus formation, and provides a means for controlling blood loss due to injury, and recruiting inflammatory cells for fighting infection. Platelet activation is not desirable however, during balloon angioplasty/stent procedures in which balloon expansion inside an artery exposes collagen, initiating thrombosis, and inflammation. We have developed biomimetic polymers, termed Peptidoglycans, composed of a dermatan sulfate backbone with covalently attached collagen-binding peptides. The peptidoglycan binds to collagen, effectively masking it from platelet activation. The lead peptidoglycan binds to collagen with high affinity (KD = 24 nm) and inhibits platelet binding and activation on collagen in both static studies and under flow, while promoting endothelial regrowth on collagen. Application for angioplasty is demonstrated in the Ossabaw miniature pig by fast delivery to the vessel wall through a therapeutic infusion catheter with a proprietary PTFE porous balloon. The peptidoglycan is an approach for locally preventing platelet deposition and activation on collagen. It can be used during angioplasty to prevent platelet deposition on target vessels and could be used in any vessel, including those not amenable to stent deployment.

  • collagen binding Peptidoglycans a biomimetic approach to modulate collagen fibrillogenesis for tissue engineering applications
    Tissue Engineering Part A, 2009
    Co-Authors: John E Paderi, Rizaldi Sistiabudi, Albena Ivanisevic, Alyssa Panitch
    Abstract:

    The small leucine-rich proteoglycans (SLRPs), prevalent in collagenous tissues, regulate collagen fibrillogenesis and provide a host of biochemical cues critical to tissue function and homeostasis. Incorporating SLRPs may enhance tissue engineering designs that mimic the native extracellular matrix, although SLRPs purified from animal sources bear low yields and lack design control. Consequently, we have designed synthetic Peptidoglycans, inspired by the native SLRP decorin, that contain a collagen-binding peptide attached to a glycosaminoglycan (GAG) chain. These Peptidoglycans modulate collagen fibrillogenesis and decrease fibril diameter in vitro, similarly to decorin, while maintaining the characteristic D-banded fibrils. Application for tissue engineering is demonstrated as these Peptidoglycans are incorporated into collagen gels seeded with smooth muscle cells. Gels formed with Peptidoglycans and decorin show a faster rate of gel compaction, and one peptidoglycan uniquely increases elastin production. The peptidoglycan design can be tailored with respect to the peptide sequence and GAG identity and is expected to have versatile application in tissue engineering.