The Experts below are selected from a list of 411 Experts worldwide ranked by ideXlab platform
Gudmundur H Gudmundsson - One of the best experts on this subject based on the ideXlab platform.
-
the expression of the gene coding for the antibacterial peptide ll 37 is induced in human keratinocytes during inflammatory disorders
Journal of Biological Chemistry, 1997Co-Authors: Margareta Frohm, Birgitta Agerberth, Ghasem Ahangari, Mona Stahlebackdahl, Sture Liden, Hans Wigzell, Gudmundur H GudmundssonAbstract:Abstract The epithelia constitute a major barrier to the environment and provide the first line of defense against invading microbes. Antimicrobial peptides are emerging as participants in the defense system of epithelial barriers in general. Originally we isolated the human antimicrobial peptide LL-37 from granulocytes. The gene (CAMP or cathelicidinantimicrobial peptide) coding for this peptide belongs to the cathelicidin family, whose members contain a conserved pro-part of the Cathelin type. The human genome seems to have only one gene of this family, whereas some mammalian species have several cathelicidin genes. In the present work we demonstrate up-regulation of this human cathelicidin gene in inflammatory skin disorders, whereas in normal skin no induction was found. By in situ hybridization and immunohistochemistry the transcript and the peptide were located in keratinocytes throughout the epidermis of the inflammatory regions. In addition, the peptide was detected in partially pure fractions derived from psoriatic scales by immunoblotting. These fractions also exhibited antibacterial activity. We propose a protective role for LL-37, when the integrity of the skin barrier is damaged, participating in the first line of defense, and preventing local infection and systemic invasion of microbes.
-
the human gene fall39 and processing of the Cathelin precursor to the antibacterial peptide ll 37 in granulocytes
FEBS Journal, 1996Co-Authors: Gudmundur H Gudmundsson, Birgitta Agerberth, Jacob Odeberg, Tomas Bergman, Berit Olsson, Rosalba SalcedoAbstract:The peptide FA-LL-37, previously termed FALL-39, was originally predicted from on ORF of a cDNA clone isolated from a human bone marrow library. This peptide was synthesized and found to have antibacterial activity. We have now characterized and sequenced the complete gene for FA-LL-37, termed FALL39. It is a compact gene of 1963 bp with four exons. Exons 1-3 code for a signal sequence and the Cathelin region. Exon 4 contains the information for the mature antibacterial peptide. Our results indicate that FALL39 is the only member of the Cathelin gene family present in the human genome. Potential binding sites for acute-phase-response factors are identified in the promoter and in intron 2. A possible role for the cytokine interleukin-6 in the regulation of FALL 39 is discussed. Anti-(FA-LL-37) IgG located the peptide in granulocytes and we isolated the mature peptide from these cells after degranulation. Structural analysis determined the mature peptide to be LL-37. To obtain LL-37 for antibacterial assays, synthetic FA-LL-37 was degraded with dipeptidyl-peptidase I. This analysis showed that mature LL-37 is a potent antibacterial peptide.
-
structure of the gene for porcine peptide antibiotic pr 39 a Cathelin gene family member comparative mapping of the locus for the human peptide antibiotic fall 39
Proceedings of the National Academy of Sciences of the United States of America, 1995Co-Authors: Gudmundur H Gudmundsson, Kristin P Magnusson, B P Chowdhary, M Johansson, Leif Andersson, Hans G BomanAbstract:Abstract PR-39 is a porcine 39-aa peptide antibiotic composed of 49% proline and 24% arginine, with an activity against Gram-negative bacteria comparable to that of tetracycline. In Escherichia coli, it inhibits DNA and protein synthesis. PR-39 was originally isolated from pig small intestine, but subsequent cDNA cloning showed that the gene is expressed in the bone marrow. The open reading frame of the clone showed that PR-39 is made as 173-aa precursor whose proregion belongs to the Cathelin family. The PR39 gene, which is rather compact and spans only 1784 bp has now been sequenced. The coding information is split into four exons. The first exon contains the signal sequence of 29 residues and the first 37 residues of the Cathelin propart. Exons 2 and 3 contain only Cathelin information, while exon 4 codes for the four C-terminal Cathelin residues and the mature PR-39 peptide extended by three residues. The sequenced upstream region (1183 bp) contains four potential recognition sites for NF-IL6 and three for APRF, transcription factors known to regulate genes for both cytokines and acute phase response factors. Genomic hybridizations revealed a fairly high level of restriction fragment length polymorphism and indicated that there are at least two copies of the PR39 gene in the pig genome. PR39 was mapped to pig chromosome 13 by linkage and in situ hybridization mapping. The gene for the human peptide antibiotic FALL-39 (also a member of the Cathelin family) was mapped to human chromosome 3, which is homologous to pig chromosome 13.
-
fall 39 a putative human peptide antibiotic is cysteine free and expressed in bone marrow and testis
Proceedings of the National Academy of Sciences of the United States of America, 1995Co-Authors: Birgitta Agerberth, Hans G Boman, Hans Gunne, Jakob Odeberg, Per Kogner, Gudmundur H GudmundssonAbstract:PR-39, a proline/arginine-rich peptide antibiotic, has been purified from pig intestine and later shown to originate in the bone marrow. Intending to isolate a clone for a human counterpart to PR-39, we synthesized a PCR probe derived from the PR-39 gene. However, when this probe was used to screen a human bone marrow cDNA library, eight clones were obtained with information for another putative human peptide antibiotic, designated FALL-39 after the first four residues. FALL-39 is a 39-residue peptide lacking cysteine and tryptophan. All human peptide antibiotics previously isolated (or predicted) belong to the defensin family and contain three disulfide bridges. The clone for prepro-FALL-39 encodes a Cathelin-like precursor protein with 170 amino acid residues. We have postulated a dibasic processing site for the mature FALL-39 and chemically synthesized the putative peptide. In basal medium E, synthetic FALL-39 was highly active against Escherichia coli and Bacillus megaterium. Residues 13-34 in FALL-39 can be predicted to form a perfect amphiphatic helix, and CD spectra showed that medium E induced 30% helix formation in FALL-39. RNA blot analyses disclosed that the gene for FALL-39 is expressed mainly in human bone marrow and testis.
Ji Ming Wang - One of the best experts on this subject based on the ideXlab platform.
-
requirement of cramp for mouse macrophages to eliminate phagocytosed e coli through an autophagy pathway
Journal of Cell Science, 2021Co-Authors: Keqiang Chen, Wanghua Gong, Teizo Yoshimura, Cuimeng Tian, Jiaqiang Huang, Giorgio Trinchieri, Ji Ming WangAbstract:Host-derived antimicrobial peptides play an important role in the defense against extracellular bacterial infections. However, the capacity of antimicrobial peptides derived from macrophages as potential antibacterial effectors against intracellular pathogens remains unknown. In this study, we report that normal (wild type, WT) mouse macrophages increased their expression of the Cathelin-related antimicrobial peptide (CRAMP) after infection by viable E. coli or stimulation with inactivated E. coli and its product LPS, a process involving activation of NF-κB followed by protease-dependent conversion of CRAMP from an inactive precursor to an active form. The active CRAMP was required by WT macrophages to eliminate phagocytosed E. coli, with participation of autophagy-related proteins ATG5, LC3-II, and LAMP-1 as well as conjugation of the bacteria with p62. This process was impaired in CRAMP -/- macrophages resulting in retention of intracellular bacteria and fragmentation of macrophages. These results indicate CRAMP as a critical component in autophagy-mediated clearance of intracellular E. coli by mouse macrophages.
-
the critical role of the antimicrobial peptide ll 37 cramp in protection of colon microbiota balance mucosal homeostasis anti inflammatory responses and resistance to carcinogenesis
Critical Reviews in Immunology, 2019Co-Authors: Meihua Zhang, Weiwei Liang, Wanghua Gong, Teizo Yoshimura, Keqiang Chen, Ji Ming WangAbstract:Mouse Cathelin-related antimicrobial peptide (CRAMP) and its homologue human cathelicidin (LL-37) play active roles in innate immune responses, angiogenesis, and wound healing. In addition, LL-37/CRAMP fends off microbes and protects against infections in the colon, where the epithelium is exposed to myriad of enteric pathogens. It is increasingly recognized that LL-37/CRAMP maintains colon mucosal barrier integrity, shapes the composition of microbiota, and protects the host from tumorigenesis. In this review, we discuss the importance of LL-37/CRAMP in the homeostasis of the host, with novel findings derived from mice deficient in CRAMP that support the proposition for this natural antimicrobial peptide and an immune modulator as a drug lead for therapeutic development.
-
the formylpeptide receptor 2 fpr2 and its endogenous ligand Cathelin related antimicrobial peptide cramp promote dendritic cell maturation
Journal of Biological Chemistry, 2014Co-Authors: Keqiang Chen, Wanghua Gong, Teizo Yoshimura, Jiaqiang Huang, Yi Xiang, Qun Jiang, Lino Tessarollo, Ji Ming WangAbstract:Mouse formylpeptide receptor 2 (Fpr2) is a homologue of the human G-protein coupled chemoattractant receptor FPR2, which interacts with pathogen and host-derived chemotactic agonists. Our previous studies revealed reduced allergic airway inflammation and immune responses in Fpr2-deficient (Fpr2(-/-)) mice in association with diminished dendritic cell (DC) recruitment into the airway and draining lymph nodes. These defects prompted us to investigate the potential changes in the differentiation and maturation of DCs caused by Fpr2 deficiency. Bone marrow monocytes from Fpr2(-/-) mouse mice incubated with GM-CSF and IL-4 in vitro showed normal expression of markers of immature DCs. However, upon stimulation with the TLR4 agonist LPS, Fpr2(-/-) mouse DCs failed to express normal levels of maturation markers with reduced production of IL-12 and diminished chemotaxis in response to the DC homing chemokine CCL21. Fpr2(-/-) DCs also failed to induce allogeneic T-cell proliferation in vitro, and their recruitment into the T-cell zones of the spleen was reduced after antigen immunization. The capacity of Fpr2 to sustain normal DC maturation was dependent on its interaction with an endogenous ligand CRAMP expressed by DCs, because neutralization of either Fpr2 or CRAMP inhibited DC maturation in response to LPS. We additionally observed that the presence of exogenous CRAMP in culture increased the sensitivity of WT mouse DCs to LPS stimulation. The importance of CRAMP for DC maturation was further demonstrated by the observations that DCs from CRAMP(-/-) mice expressed lower levels of costimulatory molecules and MHC II and exhibited poor chemotaxis in response to CCL21 after LPS stimulation. Our observations indicate a nonredundant role for Fpr2 and its agonist CRAMP in DC maturation in immune responses.
Robert I Lehrer - One of the best experts on this subject based on the ideXlab platform.
-
activities of ll 37 a Cathelin associated antimicrobial peptide of human neutrophils
Antimicrobial Agents and Chemotherapy, 1998Co-Authors: Joanne Turner, Yoon Cho, Nhunguyen Dinh, Alan J Waring, Robert I LehrerAbstract:Human neutrophils contain two structurally distinct types of antimicrobial peptides, beta-sheet defensins (HNP-1 to HNP-4) and the alpha-helical peptide LL-37. We used radial diffusion assays and an improved National Committee for Clinical Laboratory Standards-type broth microdilution assay to compare the antimicrobial properties of LL-37, HNP-1, and protegrin (PG-1). Although generally less potent than PG-1, LL-37 showed considerable activity (MIC, <10 microgram/ml) against Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli, Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, and vancomycin-resistant enterococci, even in media that contained 100 mM NaCl. Certain organisms (methicillin-resistant S. aureus, Proteus mirabilis, and Candida albicans) were resistant to LL-37 in media that contained 100 mM NaCl but were susceptible in low-salt media. Burkholderia cepacia was resistant to LL-37, PG-1, and HNP-1 in low- or high-salt media. LL-37 caused outer and inner membrane permeabilization of E. coli ML-35p. Chromogenic Limulus assays revealed that LL-37 bound to E. coli O111:B4 lipopolysaccharide (LPS) with a high affinity and that this binding showed positive cooperativity (Hill coefficient = 2.02). Circular dichroism spectrometry disclosed that LL-37 underwent conformational change in the presence of lipid A, transitioning from a random coil to an alpha-helical structure. The broad-spectrum antimicrobial properties of LL-37, its presence in neutrophils, and its inducibility in keratinocytes all suggest that this peptide and its precursor (hCAP-18) may protect skin and other tissues from bacterial intrusions and LPS-induced toxicity. The potent activity of LL-37 against P. aeruginosa, including mucoid and antibiotic-resistant strains, suggests that it or related molecules might have utility as topical bronchopulmonary microbicides in cystic fibrosis.
-
the structure of porcine protegrin genes
FEBS Letters, 1995Co-Authors: Chengquan Zhao, Tomas Ganz, Robert I LehrerAbstract:We cloned the genes of three protegrins, a family of Cathelin-associated antimicrobial peptides originally isolated from porcine leukocytes. Each gene comprised 4 exons and 3 introns, wherein Exon I encoded the signal sequence and the first 37 amino acids of Cathelin, Exons II and III contained 36 and 24 additional Cathelin residues and Exon IV contained the final two Cathelin residues followed by the protegrin sequence. This quadripartite gene structure helps explain how structurally diverse antimicrobial peptides can be expressed on common, Cathelin-containing precursors. Southern blot probed with an oligonucleotide specific for protegrin genes suggested that several identical or nearly identical protegrin genes were densely clustered in the pig chromosome.
-
the structure of porcine protegrin genes
FEBS Letters, 1995Co-Authors: Chengquan Zhao, Tomas Ganz, Robert I LehrerAbstract:We cloned the genes of three protegrins, a family of Cathelin-associated antimicrobial peptides originally isolated from porcine leukocytes. Each gene comprised 4 exons and 3 introns, wherein Exon I encoded the signal sequence and the first 37 amino acids of Cathelin, Exons II and III contained 36 and 24 additional Cathelin residues and Exon IV contained the final two Cathelin residues followed by the protegrin sequence. This quadripartite gene structure helps explain how structurally diverse antimicrobial peptides can be expressed on common, Cathelin-containing precursors. Southern blot probed with an oligonucleotide specific for protegrin genes suggested that several identical or nearly identical protegrin genes were densely clustered in the pig chromosome.
Richard L Gallo - One of the best experts on this subject based on the ideXlab platform.
-
ORIGINAL ARTICLE See related Commentary on page v Antimicrobial and Protease Inhibitory Functions of the Human
2015Co-Authors: Cathelicidin Prosequence, Mohamed Zaiou, Victor Nizet, Richard L GalloAbstract:Cathelicidins are a class of small cationic peptide anti-biotics that are expressed in skin and in other epithelial cells and are an active component of mammalian in-nate immunity. Human cathelicidin (hCAP18/LL-37) consists of a conserved prosequence called the Cathelin-like domain and a C-terminal peptide named LL-37. To date, our understanding of the Cathelin-like domain was very limited. To bring insight into the function of this evolutionarily conserved prosequence, we produced re-combinant human Cathelin-like protein and full-length hCAP18/LL-37 in Escherichia coli. As the Cathelin-like protein shares homology with the cystatin family of cy-steine protease inhibitors, we ¢rst analyzed the e¡ect of the Cathelin-like recombinant protein on the cysteine protease cathepsin L. We found that the Cathelin-like protein inhibited protease activity. Next, we tested the Cathelin-like protein for antimicrobial activity using solid phase radial di¡usion and liquid phase killing assays. The Cathelin-like prosequence, but not full-length hCAP18/LL-37, killed human pathogens including E. coli and methicillin-resistant Staphylococcus aureus at concentrations ranging from 16 to 32 lM. Together these ¢ndings suggest that after proteolytic cleavage the Cathelin-like domain can contribute to innate host defense through inhibition of bacterial growth and lim-itation of cysteine-proteinase-mediated tissue damage. As these dual functions are complementary to the LL-37 peptide released from the C-terminus of full-length hCAP18/LL-37, human cathelicidin represents an elegant multifunctional e¡ector molecule for innate im-mune defense of the skin. Key words: Innate immunity/ protease/infection/skin/staphylococcus. J Invest Dermato
-
3′ RACE Analysis and Identification of feCath.
2013Co-Authors: Brian C. Leonard, Richard L Gallo, Hiutung Chu, Jennifer L. Johns, Peter F. Moore, Stanley L. Marks, Charles L. BevinsAbstract:(A) Schematic of 3′ RACE strategy targeting the signal sequence region and the propeptide (Cathelin) domain with sense primers and using antisense adaptor primer, AP1. Feline bone marrow RNA was reverse transcribed with oligo-dT conjugated to adaptor primer 1/2 (AP1/2). Sense primers (sequences found in Table 1) were used to amplify cathelicidin related sequences in the pool of bone marrow cDNA. (B) Agarose gel electrophoresis analysis of 3′ RACE PCR products from (A). HaeIII digested Phi-X174 phage DNA used as the marker.
-
Nucleotide sequence of feCath gene and flanking region.
2013Co-Authors: Brian C. Leonard, Richard L Gallo, Hiutung Chu, Jennifer L. Johns, Peter F. Moore, Stanley L. Marks, Charles L. BevinsAbstract:(A) Gene and mRNA schematic of feCath showing four exons and interspersed intronic sequences. The mRNA sequence includes for the 5′ and 3′ untranslated regions, a coding region for signal sequence, Cathelin-like domain, mature peptide, and a polyadenylated tail. (B) Nucleotide sequence of feCath gene and flanking regions. Numbering begins arbitrarily at the 5′ most nucleotide. Lowercase letters indicate intronic and flanking sequence, uppercase letters indicate exonic sequence with deduced amino acid sequence of coding region below in three-letter code. Polyadenylation signal is underlined. The cDNA sequence was deposited in GenBank (Accession #: HQ221766).
-
Amino acid comparison of Cathelin-like domain and mature cathelicidin.
2013Co-Authors: Brian C. Leonard, Richard L Gallo, Hiutung Chu, Jennifer L. Johns, Peter F. Moore, Stanley L. Marks, Charles L. BevinsAbstract:(A) Table of sequence identity and similarity of the Cathelin-like domain from diverse groups of cathelicidins and the mature peptide of more closely related cathelicidins (PR-39 for distant comparison with mature peptides). K9CATH: dog, Bac5: cow, LL-37: human, PR-39: pig, Indolicidin: cow, mCRAMP: mouse, rCRAMP: rat. (B) Amino acid alignment of closely related mature cathelicidins. Residues in red are basic, residues in blue are acidic, and bolded residues are identical feCath sequence. pI and net charge are calculated for each peptide. Grey box indicates region of high similarity, and maintenance of cationic and hydrophobic residues. (C) Hydrophobic and polar residue clusters shown on a helical wheel projection. Based on the same analytical approach of Zelezetsky et al [25], sequence of feCath (outer ring) is compared to K9CATH (middle ring) and human LL-37 (inner ring). The analysis is divided into N- and C-terminal sequences, with residues 1–18 shown in the left wheel and 19–36 in the right wheel. Clusters of nonpolar residues are on a shaded background. Residues that deviate from the pattern of the three sequences are highlighted. The helical-breaking proline residue near the C-terminus is on a green background.
-
activation of cathepsin l by the Cathelin like domain of protegrin 3
Molecular Immunology, 2008Co-Authors: Kenshi Yamasaki, Richard L GalloAbstract:The Cathelin-like domain (CLD) of the antimicrobial cathelicidin family constitutes a unique protein family with structural similarity to cystatins, the cysteine protease inhibitors. CLDs are derived from the processed amino-terminal prosequence of the cathelicidin precursors with conservation across the vertebrate lineage ranging from fish to human. Initial attempt to characterize a possible inhibitory activity of protegrin-3 (PG3) CLD protein (a member of the multigene family of porcine cathelicidins) against several proteases led to an unexpected finding that PG3 CLD efficiently activated rather than inhibited human cathepsin L. Partial deletion of the L2 loop of PG3 CLD, a structurally equivalent region important in interaction of cystatins with proteases, significantly decreased its activating effect on cathepsin L. A complex model based on this functional loop was proposed to explain this unexpected effect, in which evolutionary emergence of completely opposite biological activity could be associated with structural discrepancies of the loop due to sequence variations between pig and human. Our results provide new insights into deeper understanding of the immune-related biological activity of this so-called pro-domain of the cathelicidin family.
Birgitta Agerberth - One of the best experts on this subject based on the ideXlab platform.
-
the expression of the gene coding for the antibacterial peptide ll 37 is induced in human keratinocytes during inflammatory disorders
Journal of Biological Chemistry, 1997Co-Authors: Margareta Frohm, Birgitta Agerberth, Ghasem Ahangari, Mona Stahlebackdahl, Sture Liden, Hans Wigzell, Gudmundur H GudmundssonAbstract:Abstract The epithelia constitute a major barrier to the environment and provide the first line of defense against invading microbes. Antimicrobial peptides are emerging as participants in the defense system of epithelial barriers in general. Originally we isolated the human antimicrobial peptide LL-37 from granulocytes. The gene (CAMP or cathelicidinantimicrobial peptide) coding for this peptide belongs to the cathelicidin family, whose members contain a conserved pro-part of the Cathelin type. The human genome seems to have only one gene of this family, whereas some mammalian species have several cathelicidin genes. In the present work we demonstrate up-regulation of this human cathelicidin gene in inflammatory skin disorders, whereas in normal skin no induction was found. By in situ hybridization and immunohistochemistry the transcript and the peptide were located in keratinocytes throughout the epidermis of the inflammatory regions. In addition, the peptide was detected in partially pure fractions derived from psoriatic scales by immunoblotting. These fractions also exhibited antibacterial activity. We propose a protective role for LL-37, when the integrity of the skin barrier is damaged, participating in the first line of defense, and preventing local infection and systemic invasion of microbes.
-
the human gene fall39 and processing of the Cathelin precursor to the antibacterial peptide ll 37 in granulocytes
FEBS Journal, 1996Co-Authors: Gudmundur H Gudmundsson, Birgitta Agerberth, Jacob Odeberg, Tomas Bergman, Berit Olsson, Rosalba SalcedoAbstract:The peptide FA-LL-37, previously termed FALL-39, was originally predicted from on ORF of a cDNA clone isolated from a human bone marrow library. This peptide was synthesized and found to have antibacterial activity. We have now characterized and sequenced the complete gene for FA-LL-37, termed FALL39. It is a compact gene of 1963 bp with four exons. Exons 1-3 code for a signal sequence and the Cathelin region. Exon 4 contains the information for the mature antibacterial peptide. Our results indicate that FALL39 is the only member of the Cathelin gene family present in the human genome. Potential binding sites for acute-phase-response factors are identified in the promoter and in intron 2. A possible role for the cytokine interleukin-6 in the regulation of FALL 39 is discussed. Anti-(FA-LL-37) IgG located the peptide in granulocytes and we isolated the mature peptide from these cells after degranulation. Structural analysis determined the mature peptide to be LL-37. To obtain LL-37 for antibacterial assays, synthetic FA-LL-37 was degraded with dipeptidyl-peptidase I. This analysis showed that mature LL-37 is a potent antibacterial peptide.
-
fall 39 a putative human peptide antibiotic is cysteine free and expressed in bone marrow and testis
Proceedings of the National Academy of Sciences of the United States of America, 1995Co-Authors: Birgitta Agerberth, Hans G Boman, Hans Gunne, Jakob Odeberg, Per Kogner, Gudmundur H GudmundssonAbstract:PR-39, a proline/arginine-rich peptide antibiotic, has been purified from pig intestine and later shown to originate in the bone marrow. Intending to isolate a clone for a human counterpart to PR-39, we synthesized a PCR probe derived from the PR-39 gene. However, when this probe was used to screen a human bone marrow cDNA library, eight clones were obtained with information for another putative human peptide antibiotic, designated FALL-39 after the first four residues. FALL-39 is a 39-residue peptide lacking cysteine and tryptophan. All human peptide antibiotics previously isolated (or predicted) belong to the defensin family and contain three disulfide bridges. The clone for prepro-FALL-39 encodes a Cathelin-like precursor protein with 170 amino acid residues. We have postulated a dibasic processing site for the mature FALL-39 and chemically synthesized the putative peptide. In basal medium E, synthetic FALL-39 was highly active against Escherichia coli and Bacillus megaterium. Residues 13-34 in FALL-39 can be predicted to form a perfect amphiphatic helix, and CD spectra showed that medium E induced 30% helix formation in FALL-39. RNA blot analyses disclosed that the gene for FALL-39 is expressed mainly in human bone marrow and testis.