Autoprocessing and self-activation of the secreted protease CPAF in Chlamydia-infected cells

Ding Chen, Lei Lei, Rhonda Flores, Zhiwei Huang, Zhongming Wu, Jijie Chai, Guangming Zhong

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The Chlamydia-secreted protease/proteasome-like activity factor (CPAF) is synthesized as a proenzyme (proCPAF) and requires processing for proteolytic activity. Recent structural studies have further demonstrated that CPAF is a serine protease that can undergo autoprocessing and self-activation in a concentration-dependent manner in vitro. However, it is not known how CPAF is processed and activated during chlamydial infection. In the current study, we used a mutant CPAF designated as CPAF(E558A) that is deficient in processing by itself as a substrate to search for putative CPAF activation factor(s) in Chlamydia-infected cells. CPAF(E558A) was processed by the lysates made from Chlamydia-infected cells and the processing activity correlated with the presence of endogenous active CPAF in the fractionated lysate samples. CPAF produced in the Chlamydia-infected cells is required for processing the mutant CPAF(E558A) since the processing activity was removed by depletion with anti-CPAF but not control antibodies. Furthermore, a purified and activated wild type CPAF alone was sufficient for processing CPAF(E558A) and no other chlamydial proteases are required. Finally, fusion tag-induced oligomerization can lead to autoprocessing and self-activation of the wild type CPAF in mammalian cells. These observations together have demonstrated that CPAF undergoes autoprocessing and self-activation during chlamydial infection.

Original languageEnglish (US)
Pages (from-to)164-173
Number of pages10
JournalMicrobial Pathogenesis
Volume49
Issue number4
DOIs
StatePublished - Oct 2010

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Chlamydia
Proteasome Endopeptidase Complex
Peptide Hydrolases
Enzyme Precursors
Serine Proteases

Keywords

  • Autoprocessing
  • Chlamydial infection
  • CPAF
  • Self-activation
  • Serine protease

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Autoprocessing and self-activation of the secreted protease CPAF in Chlamydia-infected cells. / Chen, Ding; Lei, Lei; Flores, Rhonda; Huang, Zhiwei; Wu, Zhongming; Chai, Jijie; Zhong, Guangming.

In: Microbial Pathogenesis, Vol. 49, No. 4, 10.2010, p. 164-173.

Research output: Contribution to journalArticle

Chen, Ding ; Lei, Lei ; Flores, Rhonda ; Huang, Zhiwei ; Wu, Zhongming ; Chai, Jijie ; Zhong, Guangming. / Autoprocessing and self-activation of the secreted protease CPAF in Chlamydia-infected cells. In: Microbial Pathogenesis. 2010 ; Vol. 49, No. 4. pp. 164-173.
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