Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b

E. J. Hansen, S. E. Pelzel, K. Orth, C. R. Moomaw, J. D. Radolf, C. A. Slaughter

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The P2 porin protein is the most abundant protein in the outer membrane of Haemophilus influenzae type b (Hib). Biochemical and immunochemical techniques were used to characterize the P2 proteins from a number of different Hib strains. P2 proteins from Hib outer membrane vesicles were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for in situ tryptic digestion. Solid-phase tryptic digests of P2 from eight Hib strains were resolved by high-pressure liquid chromatography and shown to be similar if not identical. Radioimmunoprecipitation analysis involving Hib cells (containing intrinsically radiolabeled proteins or lipooligosaccharide) and Western blot (immunoblot) analysis were used to identify two P2-specific murine monoclonal antibodies (MAbs). These MAbs were shown to be reactive with 120 Hib strains tested in a colony blot radioimmunoassay. One of these MAbs bound to a surface-exposed P2 epitope that was antibody accessible on all Hib strains tested; the other MAb was directed against a P2 epitope that either was not exposed on the cell surface or was otherwise inaccessible to antibody.

Original languageEnglish (US)
Pages (from-to)3270-3275
Number of pages6
JournalInfection and Immunity
Volume57
Issue number11
StatePublished - 1989

Fingerprint

Haemophilus influenzae type b
Porins
Proteins
Monoclonal Antibodies
Epitopes
Radioimmunoprecipitation Assay
Western Blotting
Collodion
Antibodies
Sodium Dodecyl Sulfate
Radioimmunoassay
Polyacrylamide Gel Electrophoresis
Digestion
High Pressure Liquid Chromatography
Membranes

ASJC Scopus subject areas

  • Immunology

Cite this

Hansen, E. J., Pelzel, S. E., Orth, K., Moomaw, C. R., Radolf, J. D., & Slaughter, C. A. (1989). Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b. Infection and Immunity, 57(11), 3270-3275.

Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b. / Hansen, E. J.; Pelzel, S. E.; Orth, K.; Moomaw, C. R.; Radolf, J. D.; Slaughter, C. A.

In: Infection and Immunity, Vol. 57, No. 11, 1989, p. 3270-3275.

Research output: Contribution to journalArticle

Hansen, EJ, Pelzel, SE, Orth, K, Moomaw, CR, Radolf, JD & Slaughter, CA 1989, 'Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b', Infection and Immunity, vol. 57, no. 11, pp. 3270-3275.
Hansen, E. J. ; Pelzel, S. E. ; Orth, K. ; Moomaw, C. R. ; Radolf, J. D. ; Slaughter, C. A. / Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b. In: Infection and Immunity. 1989 ; Vol. 57, No. 11. pp. 3270-3275.
@article{9e53b8a998f7431eb59a8508ae203278,
title = "Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b",
abstract = "The P2 porin protein is the most abundant protein in the outer membrane of Haemophilus influenzae type b (Hib). Biochemical and immunochemical techniques were used to characterize the P2 proteins from a number of different Hib strains. P2 proteins from Hib outer membrane vesicles were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for in situ tryptic digestion. Solid-phase tryptic digests of P2 from eight Hib strains were resolved by high-pressure liquid chromatography and shown to be similar if not identical. Radioimmunoprecipitation analysis involving Hib cells (containing intrinsically radiolabeled proteins or lipooligosaccharide) and Western blot (immunoblot) analysis were used to identify two P2-specific murine monoclonal antibodies (MAbs). These MAbs were shown to be reactive with 120 Hib strains tested in a colony blot radioimmunoassay. One of these MAbs bound to a surface-exposed P2 epitope that was antibody accessible on all Hib strains tested; the other MAb was directed against a P2 epitope that either was not exposed on the cell surface or was otherwise inaccessible to antibody.",
author = "Hansen, {E. J.} and Pelzel, {S. E.} and K. Orth and Moomaw, {C. R.} and Radolf, {J. D.} and Slaughter, {C. A.}",
year = "1989",
language = "English (US)",
volume = "57",
pages = "3270--3275",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Structural and antigenic conservation of the P2 porin protein among strains of Haemophilus influenzae type b

AU - Hansen, E. J.

AU - Pelzel, S. E.

AU - Orth, K.

AU - Moomaw, C. R.

AU - Radolf, J. D.

AU - Slaughter, C. A.

PY - 1989

Y1 - 1989

N2 - The P2 porin protein is the most abundant protein in the outer membrane of Haemophilus influenzae type b (Hib). Biochemical and immunochemical techniques were used to characterize the P2 proteins from a number of different Hib strains. P2 proteins from Hib outer membrane vesicles were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for in situ tryptic digestion. Solid-phase tryptic digests of P2 from eight Hib strains were resolved by high-pressure liquid chromatography and shown to be similar if not identical. Radioimmunoprecipitation analysis involving Hib cells (containing intrinsically radiolabeled proteins or lipooligosaccharide) and Western blot (immunoblot) analysis were used to identify two P2-specific murine monoclonal antibodies (MAbs). These MAbs were shown to be reactive with 120 Hib strains tested in a colony blot radioimmunoassay. One of these MAbs bound to a surface-exposed P2 epitope that was antibody accessible on all Hib strains tested; the other MAb was directed against a P2 epitope that either was not exposed on the cell surface or was otherwise inaccessible to antibody.

AB - The P2 porin protein is the most abundant protein in the outer membrane of Haemophilus influenzae type b (Hib). Biochemical and immunochemical techniques were used to characterize the P2 proteins from a number of different Hib strains. P2 proteins from Hib outer membrane vesicles were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for in situ tryptic digestion. Solid-phase tryptic digests of P2 from eight Hib strains were resolved by high-pressure liquid chromatography and shown to be similar if not identical. Radioimmunoprecipitation analysis involving Hib cells (containing intrinsically radiolabeled proteins or lipooligosaccharide) and Western blot (immunoblot) analysis were used to identify two P2-specific murine monoclonal antibodies (MAbs). These MAbs were shown to be reactive with 120 Hib strains tested in a colony blot radioimmunoassay. One of these MAbs bound to a surface-exposed P2 epitope that was antibody accessible on all Hib strains tested; the other MAb was directed against a P2 epitope that either was not exposed on the cell surface or was otherwise inaccessible to antibody.

UR - http://www.scopus.com/inward/record.url?scp=0024436194&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024436194&partnerID=8YFLogxK

M3 - Article

C2 - 2478470

AN - SCOPUS:0024436194

VL - 57

SP - 3270

EP - 3275

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 11

ER -