Lipopolysaccharides of Actinobacillus Pleuropneumoniae Bind Pig Hemoglobin

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 1995 Feb 1

PMID 7822035

Citations 15

Authors

M Belanger

C Begin

M Jacques

Affiliations

Soon will be listed here.

Abstract

A previous study indicated that lipopolysaccharides (LPS) extracted from Actinobacillus pleuropneumoniae bind two low-molecular-mass proteins, of approximately 10 and 11 kDa, present in porcine respiratory tract secretions (M. Bélanger, D. Dubreuil, and M. Jacques, Infect. Immun. 62:868-873, 1994). In the present study, we determined the N-terminal amino acid sequences of these two proteins, which revealed high homology with the alpha and beta chains of pig hemoglobin. Some isolates of A. pleuropneumoniae were able to use hemoglobin from various animal species as well as other heme compounds as sole sources of iron for growth, while other isolates were unable to use them. Immunoelectron microscopy showed binding of pig hemoglobin at the surface of all A. pleuropneumoniae isolates as well as labeling of outer membrane blebs. We observed, using Western blotting (immunoblotting), that the lipid A-core region of LPS of all isolates was binding pig hemoglobin. Furthermore, lipid A obtained after acid hydrolysis of LPS extracted from A. pleuropneumoniae was able to bind pig hemoglobin and this binding was completely abolished by preincubation of lipid A with polymyxin B but was not inhibited by preincubation with glucosamines. Fatty acids constituting the lipid A of A. pleuropneumoniae, namely, dodecanoic acid, tetradecanoic acid, 3-hydroxytetradecanoic acid, hexadecanoic acid, and octadecanoic acid, were also binding pig hemoglobin. Our results indicate that LPS of all A. pleuropneumoniae isolates tested bind pig hemoglobin and that lipid A is involved in this binding. Our results also indicate that some A. pleuropneumoniae isolates are, in addition, able to use hemoglobin for growth. Binding of hemoglobin to LPS might represent an important means by which A. pleuropneumoniae acquires iron in vivo from hemoglobin released from erythrocytes lysed by the action of its hemolysins.

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References

Wood D, Parent J, Lim E, Pruyne P, Watkins J, Spoor E . Reactivity of monoclonal antibody E5 with endotoxin. I. Binding to lipid A and rough lipopolysaccharides. Circ Shock. 1992; 38(1):55-62. View

Lynn W, Golenbock D . Lipopolysaccharide antagonists. Immunol Today. 1992; 13(7):271-6. DOI: 10.1016/0167-5699(92)90009-V. View

Roth R, LEVIN F, Levin J . Distribution of bacterial endotoxin in human and rabbit blood and effects of stroma-free hemoglobin. Infect Immun. 1993; 61(8):3209-15. PMC: 280989. DOI: 10.1128/iai.61.8.3209-3215.1993. View

Frey J, Bosse J, Chang Y, Cullen J, Fenwick B, Gerlach G . Actinobacillus pleuropneumoniae RTX-toxins: uniform designation of haemolysins, cytolysins, pleurotoxin and their genes. J Gen Microbiol. 1993; 139(8):1723-8. DOI: 10.1099/00221287-139-8-1723. View

Belanger M, Dubreuil D, Jacques M . Proteins found within porcine respiratory tract secretions bind lipopolysaccharides of Actinobacillus pleuropneumoniae. Infect Immun. 1994; 62(3):868-73. PMC: 186195. DOI: 10.1128/iai.62.3.868-873.1994. View

Rietschel E, Kirikae T, Schade F, Mamat U, Schmidt G, Loppnow H . Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB J. 1994; 8(2):217-25. DOI: 10.1096/fasebj.8.2.8119492. View

Roth R . Hemoglobin enhances the production of tissue factor by endothelial cells in response to bacterial endotoxin. Blood. 1994; 83(10):2860-5. View

Niven D, Donga J, Archibald F . Responses of Haemophilus pleuropneumoniae to iron restriction: changes in the outer membrane protein profile and the removal of iron from porcine transferrin. Mol Microbiol. 1989; 3(8):1083-9. DOI: 10.1111/j.1365-2958.1989.tb00258.x. View

Morton D, Williams P . Utilization of transferrin-bound iron by Haemophilus species of human and porcine origins. FEMS Microbiol Lett. 1989; 53(1-2):123-7. DOI: 10.1016/0378-1097(89)90378-9. View

10.

LEGENDRE N, Matsudaira P . Direct protein microsequencing from Immobilon-P Transfer Membrane. Biotechniques. 1988; 6(2):154-9. View

11.

Belanger M, Dubreuil D, Harel J, Girard C, Jacques M . Role of lipopolysaccharides in adherence of Actinobacillus pleuropneumoniae to porcine tracheal rings. Infect Immun. 1990; 58(11):3523-30. PMC: 313692. DOI: 10.1128/iai.58.11.3523-3530.1990. View

12.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

13.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

14.

Hewick R, Hunkapiller M, HOOD L, Dreyer W . A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981; 256(15):7990-7. View

15.

Tsai C, Frasch C . A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982; 119(1):115-9. DOI: 10.1016/0003-2697(82)90673-x. View

16.

Darveau R, Hancock R . Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983; 155(2):831-8. PMC: 217756. DOI: 10.1128/jb.155.2.831-838.1983. View

17.

Weinberg E . Iron withholding: a defense against infection and neoplasia. Physiol Rev. 1984; 64(1):65-102. DOI: 10.1152/physrev.1984.64.1.65. View

18.

Jensen A, Bertram T . Morphological and biochemical comparison of virulent and avirulent isolates of Haemophilus pleuropneumoniae serotype 5. Infect Immun. 1986; 51(2):419-24. PMC: 262344. DOI: 10.1128/iai.51.2.419-424.1986. View

19.

Van Lenten B, Fogelman A, Haberland M, Edwards P . The role of lipoproteins and receptor-mediated endocytosis in the transport of bacterial lipopolysaccharide. Proc Natl Acad Sci U S A. 1986; 83(8):2704-8. PMC: 323368. DOI: 10.1073/pnas.83.8.2704. View

20.

Fenwick B, Osburn B, OLANDER H . Isolation and biological characterization of two lipopolysaccharides and a capsular-enriched polysaccharide preparation from Haemophilus pleuropneumoniae. Am J Vet Res. 1986; 47(7):1433-41. View