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PMID: 1541562 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and expression of a transferrin-binding protein from Actinobacillus pleuropneumoniae.

Infection and immunity ·Vol. 60 ·No. 3 ·1992-03-00 ·Pages 892-8

Gerlach GF, Anderson C, Potter AA, Klashinsky S, Willson PJ

Abstract

An expression library was constructed from Actinobacillus pleuropneumoniae serotype 7. Escherichia coli transformants expressing recombinant proteins were identified by immunoscreening with porcine convalescent serum. One transformant expressing a 60-kDa protein (60K protein) in aggregated form was identified. Serum raised against the recombinant protein recognized a polypeptide with an indistinguishable electrophoretic mobility in the A. pleuropneumoniae wild type after iron-restricted growth only. The recombinant protein bound transferrin after blotting onto nitrocellulose. Using a competitive enzyme-linked immunosorbent assay (ELISA), the specificity of this binding for the amino-terminal half of iron-saturated porcine transferrin was established. Also, the 60K wild-type protein bound hemin as assessed by hemin-agarose chromatography. Hemin could inhibit transferrin binding of the recombinant protein in the competitive ELISA, whereas hemoglobin and synthetic iron chelators failed to do so. Southern blot analysis of several other A. pleuropneumoniae strains indicated that highly homologous sequence is present in eight of eight isolates of serotype 7 and in some isolates of serotypes 2, 3, and 4.

MeSH Terms
Actinobacillus pleuropneumoniae/chemistry,genetics Animals Carrier Proteins/analysis,genetics,physiology Cloning, Molecular Escherichia coli/genetics Genes, Bacterial Heme/metabolism Iron-Binding Proteins Swine Transferrin/metabolism Transferrin-Binding Proteins Transformation, Bacterial
Chemicals
Carrier Proteins Iron-Binding Proteins Transferrin Transferrin-Binding Proteins Heme
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gerlach G F
Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Canada.
Anderson C
Potter A A
Klashinsky S
Willson P J
References (25)
25 references, click to expand
  1. Comparative study of the iron-binding properties of human transferrins. I. Complete and sequential iron saturation and desaturation of the lactotransferrin.
    Biochim Biophys Acta. 1980 May 7;629(2):399-408 PMID: 6770907
  2. Preparation and characterization of an NH2-terminal fragment of human serum transferrin containing a single iron-binding site.
    J Biol Chem. 1980 Jan 25;255(2):708-13 PMID: 7356640
  3. Outer membranes of gram-negative bacteria. XIX. Isolation from Pseudomonas aeruginosa PAO1 and use in reconstitution and definition of the permeability barrier.
    J Bacteriol. 1978 Oct;136(1):381-90 PMID: 101518
  4. Outer membrane of Pseudomonas aeruginosa: heat- 2-mercaptoethanol-modifiable proteins.
    J Bacteriol. 1979 Dec;140(3):902-10 PMID: 118160
  5. Iron and infection.
    Microbiol Rev. 1978 Mar;42(1):45-66 PMID: 379572
  6. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  7. Isolation and molecular characterization of spontaneously occurring cytolysin-negative mutants of Actinobacillus pleuropneumoniae serotype 7.
    Infect Immun. 1991 Nov;59(11):4110-6 PMID: 1937770
  8. Molecular cloning, partial purification, and characterization of a haemin-binding lipoprotein from Haemophilus influenzae type b.
    Mol Microbiol. 1991 Feb;5(2):267-78 PMID: 2041470
  9. Expression of Neisseria meningitidis iron-regulated outer membrane proteins, including a 70-kilodalton transferrin receptor, and their potential for use as vaccines.
    Infect Immun. 1990 Sep;58(9):2875-81 PMID: 2117572
  10. Siderophore-independent acquisition of transferrin-bound iron by Haemophilus influenzae type b.
    J Gen Microbiol. 1990 May;136(5):927-33 PMID: 2143216
  11. Iron acquisition in Pasteurella haemolytica: expression and identification of a bovine-specific transferrin receptor.
    Infect Immun. 1990 Jul;58(7):2091-7 PMID: 2365453
  12. Export and intercellular transfer of DNA via membrane blebs of Neisseria gonorrhoeae.
    J Bacteriol. 1989 May;171(5):2499-505 PMID: 2496108
  13. Effect of iron restriction on the outer membrane proteins of Actinobacillus (Haemophilus) pleuropneumoniae.
    Infect Immun. 1989 Mar;57(3):798-804 PMID: 2917787
  14. Identification and characterization of the human lactoferrin-binding protein from Neisseria meningitidis.
    Infect Immun. 1988 May;56(5):1144-9 PMID: 3128478
  15. Identification and characterization of the transferrin receptor from Neisseria meningitidis.
    Mol Microbiol. 1988 Mar;2(2):281-8 PMID: 3132585
  16. Loss of transferrin receptor activity in Neisseria meningitidis correlates with inability to use transferrin as an iron source.
    Infect Immun. 1988 Dec;56(12):3132-8 PMID: 3141281
  17. Response of Neisseria gonorrhoeae to iron limitation: alterations in expression of membrane proteins without apparent siderophore production.
    Infect Immun. 1985 Feb;47(2):388-94 PMID: 3155708
  18. Haemophilus influenzae can use human transferrin as a sole source for required iron.
    Infect Immun. 1985 Apr;48(1):248-51 PMID: 3872264
  19. A simple aerosol chamber for experimental reproduction of respiratory disease in pigs and other species.
    Can J Comp Med. 1985 Oct;49(4):434-5 PMID: 3935301
  20. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  21. Iron-controlled infection with Neisseria meningitidis in mice.
    Infect Immun. 1980 Sep;29(3):886-91 PMID: 6159328
  22. Expression of a high-affinity mechanism for acquisition of transferrin iron by Neisseria meningitidis.
    Infect Immun. 1982 Apr;36(1):107-13 PMID: 6210635
  23. Iron acquisition by Neisseria meningitidis in vitro.
    Infect Immun. 1980 Feb;27(2):322-34 PMID: 6445876
  24. Enhancement of Neisseria meningitidis infection in mice by addition of iron bound to transferrin.
    Infect Immun. 1981 Oct;34(1):120-5 PMID: 6795121
  25. Construction and expression of hybrid plasmids containing the Escherichia coli glyA genes.
    Gene. 1981 Jun-Jul;14(1-2):63-72 PMID: 7021324
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.60.3.892-898.1992
Published
1992-03-00
Pages
892-8
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC257570
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.60.3.892-898.1992 PMC Full Text View journal details
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