Molecular characterization of katy (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of yersinia pestis

Emilio Garcia, Yuri A. Nedialkov, Jeffrey Elliott, Vladimir Motin, Robert R. Brubaker

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

The first temperature-dependent proteins (expressed at 37°C, but not 26°C) to be identified in Yersinia pestis were antigens 3 (fraction 1), 4 (pH 6 antigen), and 5 (hereafter termed KatY). Antigens 3 and 4 are now established virulence factors, whereas little is known about KatY, except that it is encoded chromosomally, produced in abundance, possesses modest catalase activity, and is shared by Yersinia pseudotuberculosis, but not Yersinia enterocolitica. We report here an improved chromatographic method (DEAE-cellulose, calcium hydroxylapatite, and Sephadex G-150) that yields enzymatically active KatY (2,423 U/mg of protein). Corresponding mouse monoclonal antibody 1B70.1 detected plasminogen activator-mediated hydrolysis of KatY, and a polyclonal rabbit antiserum raised against outer membranes of Y. pestis was enriched for anti-KatY. A sequenced ~16-kb Y. pestis DNA insert of a positive pLG338 clone indicated that katY encodes an 81.4-kDa protein (pI 6.98) containing a leader sequence of 2.6 kDa; the deduced molecular mass and pI of processed KatY were 78.8 kDa and 6.43, respectively. A minor truncated variant (predicted molecular mass of 53.6 kDa) was also expressed. KatY is similar (39 to 59% identity) to vegetative bacterial catalase-peroxidases (KatG in Escherichia coli) and is closely related to plasmid-encoded KatP of enterohemorrhagic E. coli O157:H7 (75% identity), katY encoded a putative Ca2+-binding site, and its promoter contained three homologues to the consensus recognition sequence of the pCD-encoded transcriptional activator LcrF. rbsA was located upstream of katY, and cybB, cybC, dmsABC, and araD were mapped downstream. These genes are not linked to katG or katP in E. coli.

Original languageEnglish (US)
Pages (from-to)3114-3122
Number of pages9
JournalJournal of Bacteriology
Volume181
Issue number10
StatePublished - May 1999
Externally publishedYes

Fingerprint

Yersinia pestis
Catalase
Peroxidase
Antigens
Yersinia pseudotuberculosis
Escherichia coli
Enterohemorrhagic Escherichia coli
DEAE-Cellulose
Yersinia enterocolitica
Proteins
Escherichia coli O157
Plasminogen Activators
Consensus Sequence
Virulence Factors
Durapatite
Immune Sera
Hydrolysis
Plasmids
Clone Cells
Binding Sites

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Molecular characterization of katy (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of yersinia pestis. / Garcia, Emilio; Nedialkov, Yuri A.; Elliott, Jeffrey; Motin, Vladimir; Brubaker, Robert R.

In: Journal of Bacteriology, Vol. 181, No. 10, 05.1999, p. 3114-3122.

Research output: Contribution to journalArticle

Garcia, Emilio ; Nedialkov, Yuri A. ; Elliott, Jeffrey ; Motin, Vladimir ; Brubaker, Robert R. / Molecular characterization of katy (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of yersinia pestis. In: Journal of Bacteriology. 1999 ; Vol. 181, No. 10. pp. 3114-3122.
@article{6fd11f80e83c4e76836e2788b949f03d,
title = "Molecular characterization of katy (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of yersinia pestis",
abstract = "The first temperature-dependent proteins (expressed at 37°C, but not 26°C) to be identified in Yersinia pestis were antigens 3 (fraction 1), 4 (pH 6 antigen), and 5 (hereafter termed KatY). Antigens 3 and 4 are now established virulence factors, whereas little is known about KatY, except that it is encoded chromosomally, produced in abundance, possesses modest catalase activity, and is shared by Yersinia pseudotuberculosis, but not Yersinia enterocolitica. We report here an improved chromatographic method (DEAE-cellulose, calcium hydroxylapatite, and Sephadex G-150) that yields enzymatically active KatY (2,423 U/mg of protein). Corresponding mouse monoclonal antibody 1B70.1 detected plasminogen activator-mediated hydrolysis of KatY, and a polyclonal rabbit antiserum raised against outer membranes of Y. pestis was enriched for anti-KatY. A sequenced ~16-kb Y. pestis DNA insert of a positive pLG338 clone indicated that katY encodes an 81.4-kDa protein (pI 6.98) containing a leader sequence of 2.6 kDa; the deduced molecular mass and pI of processed KatY were 78.8 kDa and 6.43, respectively. A minor truncated variant (predicted molecular mass of 53.6 kDa) was also expressed. KatY is similar (39 to 59{\%} identity) to vegetative bacterial catalase-peroxidases (KatG in Escherichia coli) and is closely related to plasmid-encoded KatP of enterohemorrhagic E. coli O157:H7 (75{\%} identity), katY encoded a putative Ca2+-binding site, and its promoter contained three homologues to the consensus recognition sequence of the pCD-encoded transcriptional activator LcrF. rbsA was located upstream of katY, and cybB, cybC, dmsABC, and araD were mapped downstream. These genes are not linked to katG or katP in E. coli.",
author = "Emilio Garcia and Nedialkov, {Yuri A.} and Jeffrey Elliott and Vladimir Motin and Brubaker, {Robert R.}",
year = "1999",
month = "5",
language = "English (US)",
volume = "181",
pages = "3114--3122",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "10",

}

TY - JOUR

T1 - Molecular characterization of katy (antigen 5), a thermoregulated chromosomally encoded catalase-peroxidase of yersinia pestis

AU - Garcia, Emilio

AU - Nedialkov, Yuri A.

AU - Elliott, Jeffrey

AU - Motin, Vladimir

AU - Brubaker, Robert R.

PY - 1999/5

Y1 - 1999/5

N2 - The first temperature-dependent proteins (expressed at 37°C, but not 26°C) to be identified in Yersinia pestis were antigens 3 (fraction 1), 4 (pH 6 antigen), and 5 (hereafter termed KatY). Antigens 3 and 4 are now established virulence factors, whereas little is known about KatY, except that it is encoded chromosomally, produced in abundance, possesses modest catalase activity, and is shared by Yersinia pseudotuberculosis, but not Yersinia enterocolitica. We report here an improved chromatographic method (DEAE-cellulose, calcium hydroxylapatite, and Sephadex G-150) that yields enzymatically active KatY (2,423 U/mg of protein). Corresponding mouse monoclonal antibody 1B70.1 detected plasminogen activator-mediated hydrolysis of KatY, and a polyclonal rabbit antiserum raised against outer membranes of Y. pestis was enriched for anti-KatY. A sequenced ~16-kb Y. pestis DNA insert of a positive pLG338 clone indicated that katY encodes an 81.4-kDa protein (pI 6.98) containing a leader sequence of 2.6 kDa; the deduced molecular mass and pI of processed KatY were 78.8 kDa and 6.43, respectively. A minor truncated variant (predicted molecular mass of 53.6 kDa) was also expressed. KatY is similar (39 to 59% identity) to vegetative bacterial catalase-peroxidases (KatG in Escherichia coli) and is closely related to plasmid-encoded KatP of enterohemorrhagic E. coli O157:H7 (75% identity), katY encoded a putative Ca2+-binding site, and its promoter contained three homologues to the consensus recognition sequence of the pCD-encoded transcriptional activator LcrF. rbsA was located upstream of katY, and cybB, cybC, dmsABC, and araD were mapped downstream. These genes are not linked to katG or katP in E. coli.

AB - The first temperature-dependent proteins (expressed at 37°C, but not 26°C) to be identified in Yersinia pestis were antigens 3 (fraction 1), 4 (pH 6 antigen), and 5 (hereafter termed KatY). Antigens 3 and 4 are now established virulence factors, whereas little is known about KatY, except that it is encoded chromosomally, produced in abundance, possesses modest catalase activity, and is shared by Yersinia pseudotuberculosis, but not Yersinia enterocolitica. We report here an improved chromatographic method (DEAE-cellulose, calcium hydroxylapatite, and Sephadex G-150) that yields enzymatically active KatY (2,423 U/mg of protein). Corresponding mouse monoclonal antibody 1B70.1 detected plasminogen activator-mediated hydrolysis of KatY, and a polyclonal rabbit antiserum raised against outer membranes of Y. pestis was enriched for anti-KatY. A sequenced ~16-kb Y. pestis DNA insert of a positive pLG338 clone indicated that katY encodes an 81.4-kDa protein (pI 6.98) containing a leader sequence of 2.6 kDa; the deduced molecular mass and pI of processed KatY were 78.8 kDa and 6.43, respectively. A minor truncated variant (predicted molecular mass of 53.6 kDa) was also expressed. KatY is similar (39 to 59% identity) to vegetative bacterial catalase-peroxidases (KatG in Escherichia coli) and is closely related to plasmid-encoded KatP of enterohemorrhagic E. coli O157:H7 (75% identity), katY encoded a putative Ca2+-binding site, and its promoter contained three homologues to the consensus recognition sequence of the pCD-encoded transcriptional activator LcrF. rbsA was located upstream of katY, and cybB, cybC, dmsABC, and araD were mapped downstream. These genes are not linked to katG or katP in E. coli.

UR - http://www.scopus.com/inward/record.url?scp=0032934566&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032934566&partnerID=8YFLogxK

M3 - Article

VL - 181

SP - 3114

EP - 3122

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 10

ER -