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1.
Md. Morshedur Rahman Woan-Sub Kim Toshiaki Ito Haruto Kumura Kei-ichi Shimazaki 《Anaerobe》2009,15(4):133-137
Lactoferrin, a major whey protein of human milk, is considered as growth promoter for bifidobacteria, the predominant microorganisms of human intestine. In the present study, in vitro growth promotion and cell binding ability of bovine lactoferrin to several strains of Bifidobacterium longum have been demonstrated. A dose-dependent as well as strain-dependent growth promotion effect by lactoferrin was observed. Cell binding ability of lactoferrin was inspected under an inverted confocal laser scanning microscope by incubation bacterial cells with biotinylated bovine lactoferrin and FITC-conjugated avidin. Fluorescence staining showed bovine lactoferrin binding to all tested strains. A lactoferrin-binding protein with a molecular weight of approximately 67 kDa was also detected in the extracted membrane and cytosolic fraction of each B. longum strain by far-Western blot technique using biotinylated lactoferrin and horseradish peroxidase-conjugated streptavidin. Based on these results, we suggest that existence of lactoferrin-binding protein could be a common characteristic in bifidobacteria. It can also be hypothesized that lactoferrin-binding protein in bifidobacteria is not only involved in growth stimulation mechanism but also could play different roles. 相似文献
2.
Tomasz W. Zare¸ba Corina Pascu Waleria Hryniewicz Torkel Wadström 《Current microbiology》1997,34(1):6-11
Forty-four enterococcal strains isolated from human clinical specimens were investigated for binding of 125I-labeled fibronectin, vitronectin, thrombospondin, lactoferrin, and collagen type I and IV, and for cell surface hydrophobicity.
Most strains expressed low binding of iodine-labeled human fibronectin, collagen I and IV, and higher binding of human vitronectin,
human lactoferrin, and human thrombospondin. Bacteria grown in Todd-Hewitt broth exhibited increased binding to vitronectin
and thrombospondin. In particle agglutination assays (PAA), Enterococcus faecalis strains reacted strongly with coated latex beads in contrast to E. faecium strains, which generally did not react. The ability of enterococci to bind ECM proteins was affected by heating and proteolytic
digestion, suggesting that some protein-binding components become surface exposed after treatment with proteases. The binding
of 125I-labeled proteins to E. faecalis strain E70 was inhibited when cells were preincubated with unlabeled proteins. Preincubating cells with sulfated polymers
such as dextran sulfate (M
r
5000 and 8000), pentosan sulfate and heparin decreased binding of vitronectin, lactoferrin, and thrombospondin. The binding
of lactoferrin and thrombospondin was also decreased when bacteria were preincubated with galactose, fucose, and mannosamine,
but not with mannose. All of 30 E. faecalis strains expressed pronounced surface hydrophobicity, but 10 of 14 E. faecium strains showed hydrophilic cell surface.
Received: 22 April 1996 / Accepted: 29 June 1996 相似文献
3.
Shigeo Yamamoto Noriyuki Okujo Shyoko Matsuura Ikuko Fujiwara Yasuhiro Fujita Sumio Shinoda 《Microbiology and immunology》1994,38(9):687-693
Vibrio parahaemolyticus produces a structurally novel type of siderophore, termed vibrioferrin, in response to iron-limitation. This study was performed to examine whether vibrioferrin can assimilate iron from human iron-binding proteins for growth. Comparison of the growth rates between V. parahaemolyticus AQ 3354 and its spontaneously arising, vibrioferrin-deficient mutant revealed that vibrioferrin was able to sequester iron from 30% iron-saturated human transferrin for growth, but not from human lactoferrin even if fully saturated with iron. In both strains, iron limitation induced two high-molecular-weight outer membrane proteins with apparent molecular masses of approximately 78 and 83 kDa. Since only the outer membrane fraction including these proteins showed a binding capacity to ferric vibrioferrin complex, either of them may function as its cell surface receptor. These results suggested that the organism might utilize such a source of host iron through the action of vibrioferrin during in vivo survival and proliferation, although its importance in pathogenesis is unknown. 相似文献
4.
Viejo-Díaz M Andrés MT Pérez-Gil J Sánchez M Fierro JF 《Biochemistry. Biokhimii?a》2003,68(2):217-227
A 31-amino acid synthetic peptide (NH2-FFSASCVPGADKGQFPNLCRLCAGTGENKCA-COOH) was chemically synthesized based on the amino acid sequence of a region of human lactoferrin homologous to other sequences present in the N- and C-lobes of all members of the transferrin family proteins. The peptide, termed kaliocin-1, and lactoferrin showed a bactericidal effect in assays performed in low-ionic-strength conditions. This is the first time that it is shown that the antimicrobial effect of lactoferrin depends on the extracellular cation concentration. The antimicrobial effect of kaliocin-1 was lower than that of human lactoferrin, but their activities were inhibited by Na+ or K+ in a cation concentration-dependent manner. In addition, the peptide was able to mimic native lactoferrin, inducing K+-efflux and a selective dissipation of the transmembrane electrical potential of Escherichia coli cells without causing extensive damage to the outer and inner bacterial membranes. In contrast, the peptide, but not lactoferrin, was able to permeabilize different ions through liposomal membranes. The hypothetical interaction of kaliocin-1 with a bacterial membrane compound is discussed based in the different ion flux induced on cellular and artificial membranes as well as data from circular dichroism assays. Kaliocin-1 was not cytotoxic and could be a suitable model for the design of analogs able to mimic the antibacterial effect of human lactoferrin. 相似文献
5.
Shuichi Hashizume Kazuhiko Kuroda Hiroki Murakami 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1983,763(4):377-382
A serum-free medium supplemented with growth factor(s) was devised to grow human lymphocytic cell lines. The medium was developed using human lymphocytic cell line, Bri 7 cells. In the process of constructing the medium, human lactoferrin was found to be an essential growth factor for the cell line. Human lactoferrin has higher growth stimulatory activity than human transferrin, and was sensitive to heat. Long-term cultivation of the cells was achieved in the defined medium supplemented with human lactoferrin only. The defined medium specifically supported the growth of various other human B- and T-lymphocytic cell lines but not the growth of various mouse lymphocytic cell lines. In lactoferrin-supplemented medium, the growth of some human cell lines were further stimulated by the addition of a combination of insulin, ethanolamine and selenium, or another combination of 2-mercaptoethanol and the above three factors. Bovine lactoferrin could be substituted for human lactoferrin. 相似文献
6.
Identification of lactoferrin as an essential growth factor for human lymphocytic cell lines in serum-free medium 总被引:4,自引:0,他引:4
A serum-free medium supplemented with growth factor(s) was devised to grow human lymphocytic cell lines. The medium was developed using human lymphocytic cell line, Bri 7 cells. In the process of constructing the medium, human lactoferrin was found to be an essential growth factor for the cell line. Human lactoferrin has higher growth stimulatory activity than human transferrin, and was sensitive to heat. Long-term cultivation of the cells was achieved in the defined medium supplemented with human lactoferrin only. The defined medium specifically supported the growth of various other human B- and T-lymphocytic cell lines but not the growth of various mouse lymphocytic cell lines. In lactoferrin-supplemented medium, the growth of some human cell lines were further stimulated by the addition of a combination of insulin, ethanolamine and selenium, or another combination of 2-mercaptoethanol and the above three factors. Bovine lactoferrin could be substituted for human lactoferrin. 相似文献
7.
Expression of full-length bioactive antimicrobial human lactoferrin in potato plants 总被引:22,自引:0,他引:22
A cDNA fragment encoding human lactoferrin (hLF) linked to a plant microsomal retention signal peptide (SEKDEL) was stably integrated into the Solanum tuberosum genome by Agrobacterium tumefaciens-mediated leaf disk transformation methods. The lactoferrin gene was expressed under control of both the auxin-inducible manopine synthase (mas) P2 promoter and the cauliflower mosaic virus (CaMV) 35S tandem promoter. The presence of the hLF cDNA in the genome of regenerated transformed potato plants was detected by polymerase chain reaction amplification methods. Full-length hLF protein was identified by immunoblot analysis in tuber tissue extracts from the transformed plants by immunoblot analysis. The hLF produced in transgenic plant tissues migrated during polyacrylamide gel electrophoresis as a single band with an approximate molecular mass equal to hLF. Auxin activation of the mas P2 promoter increased lactoferrin expression levels in transformed tuber and leaf tissues to approximately 0.1% of total soluble plant protein. Antimicrobial activity against four different human pathogenic bacterial strains was detected in extracts of lactoferrin-containing potato tuber tissues. This is the first report of synthesis of full length, biologically active hLF in edible plants. 相似文献
8.
Jean-Paul Perraudin Jean-Paul Prieels 《Biochimica et Biophysica Acta (BBA)/General Subjects》1982,718(1):42-48
When the cell lysis of Micrococcus luteus by hen egg white or human lysozyme is performed in the presence of bovine or human lactoferrin, a temporary increase of the turbidity of the solution as followed at 450 nm is observed. Examination of the suspension under light microscopy has proven that the protoplasts produced upon lysozyme action are agglutinated by lactoferrin. The rate of agglutination depends on pH, lactoferrin, lysozyme and cells concentrations. Agglutination is maximal at pH 5.5. Around 1.4·106 binding sites for lactoferrin per cell have been determined through a Scatchard plot analysis. The binding to the cells is not mediated by the glycosidic moiety of lactoferrin but rather by a charge-to charge interaction as succinylation of about four out of the 39 lysines of lactoferrin completely abolishes its ability to agglutinate the cells. Binding does not depend on ionic iron nor on the iron content of lactoferrin itself. 相似文献
9.
Identification of lactoferrin-binding proteins from Treponema pallidum subspecies pallidum and Treponema denticola 总被引:4,自引:1,他引:3
Teanna M. Staggs Margaret K. Greer Joel B. Baseman Stanley C. Holt Victor V. Tryon 《Molecular microbiology》1994,12(4):613-619
Lactoferrin-binding or -associated proteins were identified in Treponema pallidum subspecies pallidum and Treponema denticola by affinity column chromatography using human lactoferrin and detergent-solubilized, radiolabelled spirochaetes. Two discrete polypeptides of T. pallidum with masses of 45 and 40kDa and a broad band from 29-34 kDa exhibited association with human apo- and partially ferrated lactoferrin. T. denticola produced two proteins that associated with a lactoferrin affinity matrix (50 and 35 kDa). T. pallidum and T. denticola did not associate with soluble, human transferrin in parallel experiments. Soluble human lactoferrin competed with all lactoferrin-associated proteins from T. pallidum and T. denticola in competitive-binding assays. However, the T. denticola proteins dissociated from a lacto-ferrin-affinity matrix in the presence of differing concentrations of unlabelled, soluble lactoferrin competitor. Treatment with phospholipase D altered migration of the diffuse 29-34 kDa band of T. pallidum suggesting that the polypeptide was lipid-modified. Each of the lactoferrin-binding proteins from T. pallidum and T. denticola reacted with pooled rabbit syphilitic antisera. The lactoferrin-binding proteins of T. pallidum reacted with human sera from patients at all stages of syphilis. In addition, a monoclonal antibody generated against the 45 kDa polypeptide of T. pallidum crossreacted with the 29–34 kDa protein. 相似文献
10.
DS Coray JA Heinemann PC Tyrer JI Keenan 《World journal of microbiology & biotechnology》2012,28(5):1871-1880
Helicobacter pylori has high global infection rates and can cause other undesirable clinical manifestations such as duodenal ulcer (DU) and gastric
cancer (GC). Frequencies of re-infection after therapeutic clearance and rates of DU versus GC vary geographically and differ
markedly between developed and developing countries, which suggests additional factors may be involved. The possibility that,
in vivo, lactoferrin (Lf) may play a subtle role in modulating micronutrient availability or bacterial internalisation with
implications for disease etiology is considered. Lf is an iron binding protein produced in mammals that has antimicrobial
and immunomodulatory properties. Some bacteria that regularly colonise mammalian hosts have adapted to living in high Lf environments
and we investigated if this included the gastric pathogen H.
pylori. We found that H.
pylori was able to use iron from fully iron-saturated human Lf (hLf) whereas partially iron-saturated hLf (apo) did not increase
H.
pylori growth. Instead, apo-hLf increased adherence to and internalisation of bacteria into cultured epithelial cells. By increasing
internalisation, we speculate that apo-human lactoferrin may contribute to H.
pylori’s ability to persistence in the human stomach, an observation that potentially has implications for the risk of H.
pylori-associated disease. 相似文献
11.
Liesbeth Vogel Forien Geluk Henk Jansen Jacob Dankert Loek van Alphen 《FEMS microbiology letters》1997,156(1):165-170
Since the ability of bacteria to compete with lactoferrin for iron contributes to the pathogenesis of mucosal infections, the presence of lactoferrin receptor activity in non-encapsulated Haemophilus influenzae was investigated. The growth of 18 H. influenzae isolates from the sputum samples of chronic bronchitis patients and of six of seven H. influenzae throat isolates from healthy adults was stimulated by iron saturated human lactoferrin. Apo-lactoferrin did not stimulate the growth of H. influenzae. Human lactoferrin binding to iron limited bacteria was detected for 16 H. influenzae strains from chronic bronchitis patients and for five of seven isolates from healthy adults. We conclude that the majority of H. influenzae isolates tested bind human lactoferrin and that the iron from lactoferrin is used for growth. 相似文献
12.
Samyn-Petit B Gruber V Flahaut C Wajda-Dubos JP Farrer S Pons A Desmaizieres G Slomianny MC Theisen M Delannoy P 《Glycoconjugate journal》2001,18(7):519-527
In order to determine the N-glycosylation potential of maize, a monocotyledon expression system for the production of recombinant glycoproteins, human lactoferrin was used as a model. The human lactoferrin coding sequence was inserted into the pUC18 plasmid under control of the wheat glutenin promoter. Maize was stably transformed and recombinant lactoferrin was purified from the fourth generation seeds. Glycosylation was analysed by gas chromatography, lectin detection, glycosidase digestions and mass spectrometry. The results indicated that both N-glycosylation sites of recombinant lactoferrin are mainly substituted by typical plant paucimannose-type glycans, with 1,2-xylose and 1,3-linked fucose at the proximal N-acetylglucosamine, and that complex-type glycans with Lewisa determinants are not present in maize recombinant lactoferrin. 相似文献
13.
Human lactoferrin binds and removes the hemoglobin receptor protein of the periodontopathogen Porphyromonas gingivalis 总被引:5,自引:0,他引:5
Porphyromonas gingivalis possesses a hemoglobin receptor (HbR) protein on the cell surface as one of the major components of the hemoglobin utilization system in this periodontopathogenic bacterium. HbR is intragenically encoded by the genes of an arginine-specific cysteine proteinase (rgpA), lysine-specific cysteine proteinase (kgp), and a hemagglutinin (hagA). Here, we have demonstrated that human lactoferrin as well as hemoglobin have the abilities to bind purified HbR and the cell surface of P. gingivalis through HbR. The interaction of lactoferrin with HbR led to the release of HbR from the cell surface of P. gingivalis. This lactoferrin-mediated HbR release was inhibited by the cysteine proteinase inhibitors effective to the cysteine proteinases of P. gingivalis. P. gingivalis could not utilize lactoferrin for its growth as an iron source and, in contrast, lactoferrin inhibited the growth of the bacterium in a rich medium containing hemoglobin as the sole iron source. Lactoferricin B, a 25-amino acid-long peptide located at the N-lobe of bovine lactoferrin, caused the same effects on P. gingivalis cells as human lactoferrin, indicating that the effects of lactoferrin might be attributable to the lactoferricin region. These results suggest that lactoferrin has a bacteriostatic action on P. gingivalis by binding HbR, removing it from the cell surface, and consequently disrupting the iron uptake system from hemoglobin. 相似文献
14.
15.
Seung-Hyun Jo Suk-Yoon Kwon Doo-Sang Park Kyoung-Sil Yang Jae-Whune Kim Ki-Teak Lee Sang-Soo Kwak Haeng-Soon Lee 《Biotechnology and Bioprocess Engineering》2006,11(5):442-448
Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human,
including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption.
We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven
by an oxidative stress-inducibleSWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth
and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length
hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf)
was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration
chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of
rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was
higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by
this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification. 相似文献
16.
The effectiveness and mechanism of iron acquisition from transferrin or lactoferrin by Aeromonas hydrophila has been analyzed with regard to the pathogenesis of this microbe. The ability of A. hydrophila's siderophore, amonabactin, to remove iron from transferrin was evaluated with in vitro competition experiments. The kinetics
of iron removal from the three molecular forms of ferric transferrin (diferric, N- and C-terminal monoferric) were investigated
by separating each form by urea gel electrophoresis. The first direct determination of individual microscopic rates of iron
removal from diferric transferrin is a result. A. hydrophila 495A2 was cultured in an iron-starved defined medium and the growth monitored. Addition of transferrin or lactoferrin promoted
bacterial growth. Growth promotion was independent of the level of transferrin or lactoferrin iron saturation (between 30
and 100%), even when the protein was sequestered inside dialysis tubing. Siderophore production was also increased when transferrin
or lactoferrin was enclosed in a dialysis tube. Cell yield and growth rate were identical in experiments where transferrin
was present inside or outside the dialysis tube, indicating that binding of transferrin was not essential and that the siderophore
plays a major role in iron uptake from transferrin. The rate of iron removal from diferric transferrin shows a hyperbolic
dependence on amonabactin concentration. Surprisingly, amonabactin cannot remove iron from the more weakly binding N-terminal
site of monoferric transferrin, while it is able to remove iron from the more strongly binding C-terminal site of monoferric
transferrin. Iron from both sites is removed from diferric transferrin and it is the N-terminal site (which does not release
iron in the monoferric protein) that releases iron more rapidly! It is apparent that there is a significant interaction of
the two lobes of the protein with regard to the chelator access. Taken together, these results support an amonabactin-dependent
mechanism for iron removal by A. hydrophila from transferrin and lactoferrin. The implications of these findings for an amonabactin-dependent mechanism for iron removal
by A. hydrophila from transferrin and lactoferrin are discussed.
Received: 8 August 1999 / Accepted: 22 October 1999 相似文献
17.
The action of bovine lactoferrin saturated with iron, zinc and manganese on the intracellular multiplication of Legionella pneumophila in HeLa cells has been tested. The results obtained showed that lactoferrin did not influence the invasive efficiency of Legionella. The intracellular multiplication of the bacterium was inhibited by apo-lactoferrin and by lactoferrin saturated with manganese and zinc, whereas lactoferrin saturated with iron enhanced the intracellular growth. Experiments in parallel were performed with iron, manganese and zinc citrate to test the effect due to the metal ions alone. Even in this condition the addition of an iron chelate enhanced the multiplication of Legionella while the manganese chelate produced a certain inhibition. 相似文献
18.
Lactoferrin (Lf) is a multifunctional glycoprotein. Due to its anti-inflammatory and anti-cancer properties and the resulting therapeutical potential, lactoferrin is at present focus of a variety of research areas. The regulation of cell growth represents one of the prominent performances of lactoferrin. In this study we found lactoferrin to inhibit proliferation of the human epithelial cancer cell line HeLa. The extent of this growth inhibition was comparable to the one induced by the transforming-growth-factor-beta-1 (TGFβ1). In contrast to other cell lines where lactoferrin stimulates growth, lactoferrin failed to activate the MAP kinases ERK1/2 or p38 in HeLa cells. However, by immunocytochemistry and cell fractionation experiments, we found that lactoferrin is capable of activating the TGFβ/Smad-2 pathway. The nuclear accumulation of Smad-2 induced by Lf was comparable in magnitude to the one induced by TGFβ1. 相似文献
19.
20.
《Archives of animal nutrition》2013,67(1):10-22
Abstract Orally supplemented lactoferrin derived from bovine milk is purported to have beneficial effects on gut health of animals. Bovine lactoferrin (0, 60, or 120 mg/d) was fed to ileal cannulated, adult dogs in a replicated 3 × 3 Latin square design with 14 d periods. Control dogs tended (p = 0.06) to have higher fecal DM concentrations compared with dogs supplemented with 120 mg/d lactoferrin (34.5 vs. 32.9%). Fecal scores ranged from 3.0 – 3.3, suggesting that feces of all dogs was near the desired consistency, with dogs supplemented with 120 mg/d lactoferrin tending (p = 0.08) to have higher fecal scores. Ileal azoreductase activity tended (p < 0.10) to be higher in dogs supplemented with 60 or 120 mg/d lactoferrin (609 vs. 592 nmol/h per g ileal DM, respectively) as compared with unsupplemented dogs (272 nmol/h per g ileal DM). The following bacterial groups were measured: bifidobacteria, Campylobacter spp., Clostridium spp., eubacteria, Escherichia coli, Lactobacillus spp., Staphylococcus spp., and Streptococcus spp. Fecal streptococci concentrations were lower (p < 0.05) for dogs receiving 60 mg/d lactoferrin (8.60 log10 cfu/g fecal DM) as compared with unsupplemented dogs (9.19 log10 cfu/g fecal DM) or dogs receiving 120 mg lactoferrin/d (9.43 log10 cfu/g fecal DM). Dogs supplemented with 120 mg/d lactoferrin tended (p = 0.08) to have higher fecal indole concentrations as compared to unsupplemented dogs (1.80 vs. 1.46 μmol/g fecal DM). Because most bacterial groups measured were unaffected, it appears that lactoferrin did not exhibit prebiotic activity, and based on the data collected, lactoferrin also did not appear to have major effects on indices of health in the dog. 相似文献