Role of group A Streptococcus HtrA in the maturation of SpeB protease

Cole, JN; Aquilina, JA; Hains, PG; Henningham, A; Sriprakash, KS; Caparon, MG; Nizet, V; Kotb, M; Cordwell, SJ; Djordjevic, SP; Walker, MJ

Role of group A Streptococcus HtrA in the maturation of SpeB protease

Cole, JN Aquilina, JA Hains, PG Henningham, A Sriprakash, KS Caparon, MG Nizet, V Kotb, M Cordwell, SJ Djordjevic, SP

Walker, MJ

Permalink

Publication Type:: Journal Article
Citation:: Proteomics, 2007, 7 (24), pp. 4488 - 4498
Issue Date:: 2007-12-01

Closed Access

	Filename	Description	Size
	2008007074OK.pdf		1.02 MB		View/Open

Copyright Clearance Process

Recently Added
In Progress
Closed Access

This item is closed access and not available.

Full metadata record

Field	Value	Language
dc.contributor.author	Cole, JN	en_US
dc.contributor.author	Aquilina, JA	en_US
dc.contributor.author	Hains, PG	en_US
dc.contributor.author	Henningham, A	en_US
dc.contributor.author	Sriprakash, KS	en_US
dc.contributor.author	Caparon, MG	en_US
dc.contributor.author	Nizet, V	en_US
dc.contributor.author	Kotb, M	en_US
dc.contributor.author	Cordwell, SJ	en_US
dc.contributor.author	Djordjevic, SP https://orcid.org/0000-0001-9301-5372	en_US
dc.contributor.author	Walker, MJ	en_US
dc.date.issued	2007-12-01	en_US
dc.identifier.citation	Proteomics, 2007, 7 (24), pp. 4488 - 4498	en_US
dc.identifier.issn	1615-9853	en_US
dc.identifier.uri	http://hdl.handle.net/10453/8666
dc.description.abstract	The serine protease high-temperature requirement A (HtrA) (DegP) of the human pathogen Streptococcus pyogenes (group A Streptococcus; GAS) is localized to the ExPortal secretory microdomain and is reportedly essential for the maturation of cysteine protease streptococcal pyrogenic exotoxin B (SpeB). Here, we utilize HSC5 (M5 serotype) and the in-frame isogenic mutant HSC5ΔhtrA to determine whether HtrA contributes to the maturation of other GAS virulence determinants. Mutanolysin cell wall extracts and secreted proteins were arrayed by 2-DE and identified by MALDI-TOF PMF analysis. HSC5ΔhtrA had elevated levels of cell wall-associated M protein, whilst the supernatant had higher concentrations of M protein fragments and a reduced amount of mature SpeB protease, compared to wild-type (WT). Western blot analysis and protease assays revealed a delay in the maturation of SpeB in the HSC5ΔhtrA supernatant. HtrA was unable to directly process SpeB zymogen (proSpeB) to the active form in vitro. We therefore conclude that HtrA plays an indirect role in the maturation of cysteine protease SpeB. © 2007 Wiley-VCH Verlag GmbH & Co. KGaA.	en_US
dc.relation.ispartof	Proteomics	en_US
dc.relation.isbasedon	10.1002/pmic.200700626	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Cell Wall	en_US
dc.subject.mesh	Streptococcus pyogenes	en_US
dc.subject.mesh	Enzyme Precursors	en_US
dc.subject.mesh	Cysteine Endopeptidases	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Proteome	en_US
dc.subject.mesh	Culture Media	en_US
dc.subject.mesh	Electrophoresis, Gel, Two-Dimensional	en_US
dc.subject.mesh	Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization	en_US
dc.subject.mesh	Enzyme Activation	en_US
dc.subject.mesh	Kinetics	en_US
dc.subject.mesh	Time Factors	en_US
dc.title	Role of group A Streptococcus HtrA in the maturation of SpeB protease	en_US
dc.type	Journal Article
utslib.citation.volume	24	en_US
utslib.citation.volume	7	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	08 Information and Computing Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	24	en_US
pubs.publication-status	Published	en_US
pubs.volume	7	en_US

Abstract:

The serine protease high-temperature requirement A (HtrA) (DegP) of the human pathogen Streptococcus pyogenes (group A Streptococcus; GAS) is localized to the ExPortal secretory microdomain and is reportedly essential for the maturation of cysteine protease streptococcal pyrogenic exotoxin B (SpeB). Here, we utilize HSC5 (M5 serotype) and the in-frame isogenic mutant HSC5ΔhtrA to determine whether HtrA contributes to the maturation of other GAS virulence determinants. Mutanolysin cell wall extracts and secreted proteins were arrayed by 2-DE and identified by MALDI-TOF PMF analysis. HSC5ΔhtrA had elevated levels of cell wall-associated M protein, whilst the supernatant had higher concentrations of M protein fragments and a reduced amount of mature SpeB protease, compared to wild-type (WT). Western blot analysis and protease assays revealed a delay in the maturation of SpeB in the HSC5ΔhtrA supernatant. HtrA was unable to directly process SpeB zymogen (proSpeB) to the active form in vitro. We therefore conclude that HtrA plays an indirect role in the maturation of cysteine protease SpeB. © 2007 Wiley-VCH Verlag GmbH & Co. KGaA.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/8666