Characterization of an OmpA-like outer membrane protein of the acidophilic iron-oxidizing bacterium, Acidithiobacillus ferrooxidans

Mohammed Abul Manchur, Mei Kikumoto, Tadayoshi Kanao, Jun Takada, Kazuo Kamimura

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

An OmpA family protein (FopA) previously reported as one of the major outer membrane proteins of an acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans was characterized with emphasis on the modification by heat and the interaction with peptidoglycan. A 30-kDa band corresponding to the FopA protein was detected in outer membrane proteins extracted at 75°C or heated to 100°C for 10 min prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). However, the band was not detected in outer membrane proteins extracted at ≤40°C and without boiling prior to electrophoresis. By Western blot analysis using the polyclonal antibody against the recombinant FopA, FopA was detected as bands with apparent molecular masses of 30 and 90 kDa, suggesting that FopA existed as an oligomeric form in the outer membrane of A. ferrooxidans. Although the fopA gene with a sequence encoding the signal peptide was successfully expressed in the outer membrane of Escherichia coli, the recombinant FopA existed as a monomer in the outer membrane of E. coli. FopA was detected in peptidoglycan-associated proteins from A. ferrooxidans. The recombinant FopA also showed the peptidoglycan-binding activity.

Original languageEnglish
Pages (from-to)403-410
Number of pages8
JournalExtremophiles
Volume15
Issue number3
DOIs
Publication statusPublished - May 2011

Fingerprint

Acidithiobacillus
Peptidoglycan
Membrane Proteins
Iron
Bacteria
Membranes
Escherichia coli
Staphylococcal Protein A
Protein Sorting Signals
Sodium Dodecyl Sulfate
Electrophoresis
Polyacrylamide Gel Electrophoresis
Hot Temperature
Western Blotting
Antibodies
Genes
Proteins

Keywords

  • Acidithiobacillus ferrooxidans
  • Acidophile
  • Iron-oxidizing bacterium
  • OmpA
  • Outer membrane protein

ASJC Scopus subject areas

  • Molecular Medicine
  • Microbiology

Cite this

Characterization of an OmpA-like outer membrane protein of the acidophilic iron-oxidizing bacterium, Acidithiobacillus ferrooxidans. / Manchur, Mohammed Abul; Kikumoto, Mei; Kanao, Tadayoshi; Takada, Jun; Kamimura, Kazuo.

In: Extremophiles, Vol. 15, No. 3, 05.2011, p. 403-410.

Research output: Contribution to journalArticle

@article{15d593833ee045f5aba529800ef0fa45,
title = "Characterization of an OmpA-like outer membrane protein of the acidophilic iron-oxidizing bacterium, Acidithiobacillus ferrooxidans",
abstract = "An OmpA family protein (FopA) previously reported as one of the major outer membrane proteins of an acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans was characterized with emphasis on the modification by heat and the interaction with peptidoglycan. A 30-kDa band corresponding to the FopA protein was detected in outer membrane proteins extracted at 75°C or heated to 100°C for 10 min prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). However, the band was not detected in outer membrane proteins extracted at ≤40°C and without boiling prior to electrophoresis. By Western blot analysis using the polyclonal antibody against the recombinant FopA, FopA was detected as bands with apparent molecular masses of 30 and 90 kDa, suggesting that FopA existed as an oligomeric form in the outer membrane of A. ferrooxidans. Although the fopA gene with a sequence encoding the signal peptide was successfully expressed in the outer membrane of Escherichia coli, the recombinant FopA existed as a monomer in the outer membrane of E. coli. FopA was detected in peptidoglycan-associated proteins from A. ferrooxidans. The recombinant FopA also showed the peptidoglycan-binding activity.",
keywords = "Acidithiobacillus ferrooxidans, Acidophile, Iron-oxidizing bacterium, OmpA, Outer membrane protein",
author = "Manchur, {Mohammed Abul} and Mei Kikumoto and Tadayoshi Kanao and Jun Takada and Kazuo Kamimura",
year = "2011",
month = "5",
doi = "10.1007/s00792-011-0371-6",
language = "English",
volume = "15",
pages = "403--410",
journal = "Extremophiles : life under extreme conditions",
issn = "1431-0651",
publisher = "Springer Japan",
number = "3",

}

TY - JOUR

T1 - Characterization of an OmpA-like outer membrane protein of the acidophilic iron-oxidizing bacterium, Acidithiobacillus ferrooxidans

AU - Manchur, Mohammed Abul

AU - Kikumoto, Mei

AU - Kanao, Tadayoshi

AU - Takada, Jun

AU - Kamimura, Kazuo

PY - 2011/5

Y1 - 2011/5

N2 - An OmpA family protein (FopA) previously reported as one of the major outer membrane proteins of an acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans was characterized with emphasis on the modification by heat and the interaction with peptidoglycan. A 30-kDa band corresponding to the FopA protein was detected in outer membrane proteins extracted at 75°C or heated to 100°C for 10 min prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). However, the band was not detected in outer membrane proteins extracted at ≤40°C and without boiling prior to electrophoresis. By Western blot analysis using the polyclonal antibody against the recombinant FopA, FopA was detected as bands with apparent molecular masses of 30 and 90 kDa, suggesting that FopA existed as an oligomeric form in the outer membrane of A. ferrooxidans. Although the fopA gene with a sequence encoding the signal peptide was successfully expressed in the outer membrane of Escherichia coli, the recombinant FopA existed as a monomer in the outer membrane of E. coli. FopA was detected in peptidoglycan-associated proteins from A. ferrooxidans. The recombinant FopA also showed the peptidoglycan-binding activity.

AB - An OmpA family protein (FopA) previously reported as one of the major outer membrane proteins of an acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans was characterized with emphasis on the modification by heat and the interaction with peptidoglycan. A 30-kDa band corresponding to the FopA protein was detected in outer membrane proteins extracted at 75°C or heated to 100°C for 10 min prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). However, the band was not detected in outer membrane proteins extracted at ≤40°C and without boiling prior to electrophoresis. By Western blot analysis using the polyclonal antibody against the recombinant FopA, FopA was detected as bands with apparent molecular masses of 30 and 90 kDa, suggesting that FopA existed as an oligomeric form in the outer membrane of A. ferrooxidans. Although the fopA gene with a sequence encoding the signal peptide was successfully expressed in the outer membrane of Escherichia coli, the recombinant FopA existed as a monomer in the outer membrane of E. coli. FopA was detected in peptidoglycan-associated proteins from A. ferrooxidans. The recombinant FopA also showed the peptidoglycan-binding activity.

KW - Acidithiobacillus ferrooxidans

KW - Acidophile

KW - Iron-oxidizing bacterium

KW - OmpA

KW - Outer membrane protein

UR - http://www.scopus.com/inward/record.url?scp=79955523356&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955523356&partnerID=8YFLogxK

U2 - 10.1007/s00792-011-0371-6

DO - 10.1007/s00792-011-0371-6

M3 - Article

VL - 15

SP - 403

EP - 410

JO - Extremophiles : life under extreme conditions

JF - Extremophiles : life under extreme conditions

SN - 1431-0651

IS - 3

ER -